Histometric Analysis Research Articles

Tissue response in a rat model of denture stomatitis treated with tissue conditioner containing antifungal complexed with β-cyclodextrin.

Introduction. Tissue conditioners modified with antifungals are a potential alternative to denture stomatitis (DS) treatment.Gap Statement. Information on tissue response to this treatment before its clinical application is lacking.Aim. This study aimed to evaluate the tissue response of a tissue conditioner modified with antifungals in a rat model of DS.Methodology. After DS induction for 4 days under antibiotic therapy, Wistar rats had their intraoral devices (IODs) relined with the tissue conditioner Softone without (Soft) or with the MICs against Candida albicans of nystatin (Nys) or chlorhexidine (Chx) complexed or not with β-cyclodextrin (Nys:βCD and Chx:βCD). Three controls were included: healthy rats [negative control (Nc)], rats using a sterile IOD [sterile device (Sd)] and rats with DS that did not receive treatment (DS). After 4 days of treatment, the palatal mucosa under the IODs underwent histological processing for morphohistopathological and histometric analyses, morphology of collagen fibres (birefringence), immunohistochemistry for the expression of cell proliferation (proliferating cell nuclear antigen) and cytokine (IL-1β).Results. The Nc and Sd groups were similar (P>0.05), displaying epithelial and connective tissues without any discernible changes in the parameters assessed. The DS and Soft groups exhibited pronounced epithelial alterations, cell proliferation and expression of the cytokine IL-1β. In groups treated with drug incorporation (Nys, Chx, Nys:βCD and Chx:βCD), all samples demonstrated a reduction in tissue inflammation or complete tissue recovery, with an epithelium compatible with health. For the immunohistochemical parameters, the Chx, Nys:βCD and Chx:βCD groups were comparable with Nc (P>0.05).Conclusion. The proposed treatment could be promising for DS, as it led to the tissue recovery of the palatal mucosa. Nevertheless, much lower concentrations of complexed antifungals were required to achieve a similar or higher degree of tissue response compared with uncomplexed drugs in a modified tissue conditioner formulation.

Aquamimicry feeding system improved growth, physiological status, muscle and intestine histomorphology, and flesh quality of keeled mullet, Liza carinata

The aquamimicry feeding system (AMFS) is one of the modern aquaculture systems that aim to reduce feed costs and water exchange rates to improve the sustainability of aquaculture projects. In AMFS, the aquatic environment is simulated to produce natural food by adding probiotics to carbon sources. Therefore, a 108-day field study was conducted to evaluate the efficiency of AMFS as a solo feed or in combination with a commercial diet for the rearing of keeled mullet, Liza carinata. The L. carinata fingerlings with an initial body weight of 30.0 ± 3.25 g were randomly distributed into three experimental treatments in triplicate rectangular polyethylene ponds (measured 7 × 3 × 1 m) at an initial stocking density of 105 fish per pond. The fish were fed one of the following regimes: the 1st is a commercial floating diet (CFD, 30 % crude protein; T1), the 2nd is a combination of 50 % CFD: 50 % AMFS (T2), the 3rd is a solo of AMFS (T3). Water quality parameters did not differ significantly among different treatments except for a significant reduction in total ammonia nitrogen and a higher zooplankton count in the water of the AMFS group. The fish reared with AMFS as a solo feed (T3) experienced a significant growth performance, somatic indexes, and digestive enzyme activities compared to the fish reared in other treatments. Fish fed AMFS alone (T3) or in combination with CFD (T2) had an improved hematological profile, liver and kidney function indicators, serum lipid, and protein profiles than those fed CFD (T1). Histological and histometric analysis of the intestine showed an improved architecture with AMFS feeding. Furthermore, AMFS feeding increased the diameter of muscle bundles and the protein content of the dorsal muscles. The flesh quality parameters, including water holding capacity, stored loss, drip loss, and frozen leakage rate were improved in fish fed on AMFS alone or in combination with CFD. Therefore, from the above-mentioned beneficial effects of AMFS, it could be used as a promising, economical, and sustainable technology for rearing L. carinata.

The Role of Chitosan and Gelatin-Based Scaffolds in Bone Regeneration: A Systematic Review.

Guided bone regeneration facilitates the growth of new bone in areas where there is a bone defect or insufficiency. This technique involves placing a barrier membrane over the bone graft site, and the membrane prevents the invasion of soft tissue (such as gingival tissue) into the bone graft area. This allows the slower-growing bone cells to populate the area without competition, promoting proper bone regeneration. When combined, chitosan and gelatin create composite scaffolds that leverage the strengths of both materials. Chitosan provides structural integrity and antimicrobial properties, and gelatin enhances cell attachment and proliferation, which improves mechanical properties and makes it more suitable for supporting bone regeneration in load-bearing areas.This systematic review aims to evaluate the effectiveness ofchitosan and gelatin-based scaffolds in bone regeneration. Various databases such as PubMed, Cochrane Library, LILAC, and Google Scholar were screened to adhere to the eligibility criteria. The included studies in the review were the in vivo and in vitro assessment of the chitosan and gelatin efficiency as a scaffold. Six studies were investigated for the involvement of chitosan and gelatin-based scaffolds in bone regeneration. Of these, two in vivostudies examined bone regeneration by measuring alkaline phosphatase activity (ALP) using different staining techniques, while the remaining four in vitro studies used histologic and histometric analysis where stem cells, chemicals, and other biopolymers were compared. Chitosan and gelatin scaffolds consistently showed better results in terms of bone repair throughout all six experiments. Gelatin's capacity for regeneration can be increased by mixing itwith chitosan. For additional advancement, future researchers need to focus on incorporating biopolymers.The potential of scaffolds composed of gelatin and chitosan to replace tissue lost due to periodontitis shows great clinical significance.

Early Peri-Implant Bone Healing on Laser-Modified Surfaces with and without Hydroxyapatite Coating: An In Vivo Study.

(1) Objective: The aim of this study was to assess the biological behavior of bone tissue on a machined surface (MS) and modifications made by a laser beam (LS) and by a laser beam incorporated with hydroxyapatite (HA) using a biomimetic method without thermic treatment (LHS). (2) Methods: Scanning electron microscopy coupled with energy-dispersive X-ray spectrometry (SEM/EDX) was performed before and after installation in the rabbit tibiae. A total of 20 Albinus rabbits randomly received 30 implants of 3.75 × 10 mm in the right and left tibias, with two implants on each surface in each tibia. In the animals belonging to the 4-week euthanasia period group, intramuscular application of the fluorochromes calcein and alizarin was performed. In implants placed mesially in the tibiofemoral joint, biomechanical analysis was performed by means of a removal torque (N/cm). The tibias with the implants located distally to the joint were submitted for analysis by confocal laser microscopy (mineral apposition rate) and for histometric analysis by bone contact implant (%BIC) and newly formed bone area (%NBA). (3) Results: The SEM showed differences between the surfaces. The biomechanical analysis revealed significant differences in removal torque values between the MSs and LHSs over a 2-week period. Over a 4-week period, both the LSs and LHSs demonstrated removal torque values statistically higher than the MSs. BIC of the LHS implants were statistically superior to MS at the 2-week period and LHS and LS surfaces were statistically superior to MS at the 4-week period. Statistical analysis of the NBA of the implants showed difference between the LHS and MS in the period of 2 weeks. (4) Conclusions: The modifications of the LSs and LHSs provided important physicochemical modifications that favored the deposition of bone tissue on the surface of the implants.

Efficacy of plasma treatment for surface cleansing and osseointegration of sandblasted and acid-etched titanium implants.

This study was conducted to evaluate the effects of plasma treatment of sandblasted and acid-etched (SLA) titanium implants on surface cleansing and osseointegration in a beagle model. For morphological analysis and XPS analysis, scanning electron microscope and x-ray photoelectron spectroscopy were used to analyze the surface topography and chemical compositions of implant before and after plasma treatment. For this animal experiment, twelve SLA titanium implants were divided into two groups: a control group (untreated implants) and a plasma group (implants treated with plasma). Each group was randomly located in the mandibular bone of the beagle dog (n = 6). After 8 weeks, the beagle dogs were sacrificed, and volumetric analysis and histometric analysis were performed within the region of interest. In morphological analysis, plasma treatment did not alter the implant surface topography or cause any physical damage. In XPS analysis, the atomic percentage of carbon at the inspection point before the plasma treatment was 34.09%. After the plasma treatment, it was reduced to 18.74%, indicating a 45% reduction in carbon. In volumetric analysis and histometric analysis, the plasma group exhibited relatively higher mean values for new bone volume (NBV), bone to implant contact (BIC), and inter-thread bone density (ITBD) compared to the control group. However, there was no significant difference between the two groups (P > .05). Within the limits of this study, plasma treatment effectively eliminated hydrocarbons without changing the implant surface.

Histological evaluation of osseointegration between conventional and novel bone-level tapered implants in healed bone-A preclinical study.

To histologically compare osseointegration and crestal bone healing between newly introduced tapered, self-cutting bone-level test implants and tapered bone-level control implants in sites with fully healed sites. Sixty-six implants (33 test, 33 control) were placed 1 mm subcrestally in a minipig model and underwent qualitative histologic and quantitative histometric analyses after 3, 6 and 12 weeks of submerged healing. The primary and secondary outcomes were the bone-to-implant contact (BIC) and first bone-to-implant contact (fBIC). Outcomes between the test and control implants were statistically compared. The BIC values of the test implants were comparable and non-inferior over the time points studied, except for the 12 weeks time point which showed statistically significantly higher BIC values of the test (88.07 ± 5.35%) compared to the control implants (80.88 ± 7.51%) (p = .010). Similarly comparable and non-inferior were the fBIC values, except for the 6-week outcome, which showed statistically higher values for the test (-546.5 ± 450.80 μm) compared to the control implants (-75.7 ± 100.59 μm). fBIC results for the test implants were qualitatively more stable and consistent between test time points. Novel self-cutting bone-level test implants demonstrated superior osseointegration and similar bone levels compared to conventional bone-level implants after a healing period of 12 weeks in healed ridges.

Combination of a Synthetic Bioceramic Associated with a Polydioxanone-Based Membrane as an Alternative to Autogenous Bone Grafting.

The purpose of this study was to evaluate the repair process in rat calvaria filled with synthetic biphasic bioceramics (Plenum® Osshp-70:30, HA:βTCP) or autogenous bone, covered with a polydioxanone membrane (PDO). A total of 48 rats were divided into two groups (n = 24): particulate autogenous bone + Plenum® Guide (AUTOPT+PG) or Plenum® Osshp + Plenum® Guide (PO+PG). A defect was created in the calvaria, filled with the grafts, and covered with a PDO membrane, and euthanasia took place at 7, 30, and 60 days. Micro-CT showed no statistical difference between the groups, but there was an increase in bone volume (56.26%), the number of trabeculae (2.76 mm), and intersection surface (26.76 mm2) and a decrease in total porosity (43.79%) in the PO+PG group, as well as higher values for the daily mineral apposition rate (7.16 µm/day). Histometric analysis presented material replacement and increased bone formation at 30 days compared to 7 days in both groups. Immunostaining showed a similar pattern between the groups, with an increase in proteins related to bone remodeling and formation. In conclusion, Plenum® Osshp + Plenum® Guide showed similar and sometimes superior results when compared to autogenous bone, making it a competent option as a bone substitute.

MORPHOLOGICAL CHANGES OF RAT PERIODONTIUM IN SUPRAOCCLUSAL RELATIONS OF INDIVIDUAL TEETH IN THE AGE ASPECT

Although the issue of the occlusal load influence on the occurrence of pathological phenomena in the body has been discussed for decades, it is still a subject of debate. Pathogenic occlusion is defined as excessive occlusal forces (premature contacts and interference) that can cause injuries to the dentition, including teeth and periodontal tissues. Changes in occlusal dimensions, if not properly performed or treated, can lead to dysfunctions of the dentition, such as joint clicking, difficulty in chewing, occlusal discomfort, muscle fatigue, and pain.
 Aim: to determine the morphological changes of periodontium in rats of different age groups in the supraocclusal relationship of individual teeth in the age aspect.
 Materials and methods of the study. The study was conducted on 60 white laboratory rats aged 4 to 22 months and weighing 90 to 330 g. The rats were divided into control (30 animals) and experimental (30 animals) groups. Each group was divided into 3 subgroups (10 animals each) depending on the age of the rats: young, mature and senile. Young animals included rats aged 3 4 months with a weight of 90 100 g; mature animals aged 6-8 months with a weight of 150 170 g; senile rats 20 22 months and weighing 200 220 g. In the experimental group, the state of supraocclusion was modeled by increasing the height of the lower right second molars by placing 1 mm high filling. For the placement of the filling, 37% orthophosphorus etching gel for enamel and dentin (Latus, Ukraine), microapplicators (Latus, Ukraine), the fifth generation Prime&Bond NT adhesive system (Dentsply Sirona, Germany), the universal microhybrid light-curing composite material Latelux (Latus, Ukraine) and the photopolymerizer LED.D (WOODPECKER, China) were used according to the manufacturer's instructions. The experimental animals were withdrawn from the experiment on the 15th day by decapitation under general anesthesia with sodium thiopental. The lower jaws were skeletonized, and dental blocks were cut out in the area of the molars. For histological examination, the dental blocks were fixed in a 10% solution of neutral formalin for two days, dehydrated in alcohols of increasing concentration and embedded in paraffin. Then, sections were made from the obtained preparations on a Shandon Finesse 325 rotary microtome with a section thickness of 4-6 μm. The histological sections were fixed on degreased slides and dried. Van Gieson staining was performed. The width of the periodontal gap was determined using the ZEISS ZEN 3.7 program by Carl Zeiss Microscopy.
 Results of the study. Histometric analysis has showed significant morphological changes in the experimental group of all age categories. In young rats, the fibrinoid edema phenomena of the periodontal ligament collagen fibers were observed. In mature rats, there was a picture of mucoid edema of collagen fibers of the periodontal ligament. In senile rats, fibrinoid necrosis of collagen fibers of the periodontal ligament were discovered, an inflammatory infiltrate, macrophages and neutrophils, with a predominance of neutrophilic inflammatory microenvironment, and initial hyaline tissue changes were found. In all age groups, there was cement resorption on the lateral surfaces of the tooth root, lacunar resorption of alveolar bone tissue and the presence of a large number of osteoclasts. In senile rats, hypercementosis was determined in the apical part of the root. In the control group of all age categories, neither resorption nor disorganization of connective tissue, nor hypercementosis was detected.
 Conclusions. The study shows that traumatic occlusion leads to resorption of root and alveolar cementum in all age groups. In young rats, traumatic occlusion causes fibrinoid edema of collagen fibers of the periodontal ligament. In mature rats, traumatic occlusion causes mucoid edema of collagen fibers of the periodontal ligament. In senile rats, traumatic occlusion causes fibrinoid necrosis of collagen fibers of the periodontal ligament, initial hyaline tissue changes and hypercementosis.

Phytochemical analysis underlying membrane stabilization and anti-oxidant promising potentials of Acacia nilotica seed extract

This study has aimed to evaluate the Acacia nilotica seed extract as a natural antioxidant and membrane stabilizer, as well as screening phytochemicals contained in the extract that may underline these activities. Antioxidant activity was evaluated using Phospho-molybdenum and DPPH antioxidant assays; membrane stabilization was evaluated using HRBC-MS assay, whereas phytochemicals were detected using qualitative analytical tests. Acacia nilotica hydro-methanolic extract exhibited potent, concentration (4-512 μg/mL)-dependent antioxidant activity indicated by Phospho-molybdenum and DPPH antioxidant assays, with IC50 values of 100.5 and 33.19 μg/mL, respectively. In addition, the extract exerted membrane stabilizing effects in a concentration (4-512 μg/mL)-dependent manner, with an IC50 value of 41.47 μg/mL. Phytochemical analysis of the plant extract clarified the active biological constituent(s) underlying these effects, including tannins, saponins, gums, flavonoids, anthraquinone glycosides, carbohydrates and proteins. These data may suggest Acacia nilotica extract, according to in vitro assays, as a highly potent antioxidant and membrane stabilizer derived from nature and could be applied as an adjunct therapy in disease conditions associated with inflammation and oxidative stress. Keywords: Acacia nilotica; antioxidant; membrane stabilizer; anti-inflammatory.

Evaluating the protective effect of alpha-lipoic acid against gentamicin-induced gonadal toxicity indicated by histopathology.

Gentamicin induces gonadotoxicity in animal models, accompanied by oxidative stress. This study evaluated the histopathological protective effect of alpha-lipoic acid against gentamicin-induced gonadotoxicity. A parallel experimental study included 50 albino Wister rats, which were divided into 5 groups of ten according to the intraperitoneal intervention: group I received gentamicin, group II received gentamicin plus alpha-lipoic (100 mg/kg), group III received gentamicin plus a double dose of alpha-lipoic acid, group III received gentamicin plus oral vitamin E, and group IV received NaCl 0.9% (control). The animals were equally euthanized on days 15 and 16. Tests were dissected, prepared, and stained using a light microscope for histopathological examination. Rats exposed to gentamicin showed degeneration of seminiferous tubules, characterized by a significant decrease in germinal epithelial cells, impaired spermatogenesis, and a lack of spermatozoa in the lumen. An improvement in testes of animals co-treated with alpha-lipoic acid or vitamin E, including restoration of spermatogenesis and epithelial thickness. The histometric analysis also showed that the tubule epithelial thickness decreased when gentamicin was used, but this did not happen when alpha-lipoic acid or vitamin E was added simultaneously. The detrimental effects of gentamicin on testicular architecture are preventable through alpha-lipoic acid or vitamin E co-treatment. Keywords: Alpha-lipoic acid; Gonads; Histometric; Gentamicin; Seminiferous tubule; Spermatogenesis.

Influence of systemic antibiotic therapy on the development and progression of induced apical periodontitis in Wistar rats.

The aim of this study was to investigate the influence of systemic antibiotic therapy on the development and progression of induced apical periodontitis (AP) in Wistar rats. Fifty-six rats were submitted to pulp exposure of the lower left first molar for the induction of AP. On the same day, intraperitoneal antibiotic therapy was administered once a day, for 15days, until euthanasia. The groups were formed according to the different treatments (n = 8): C-control; GEN-treated with gentamicin (10mg/Kg); AC-treated with amoxicillin (100mg/Kg); MZ-treated with metronidazole (40mg/Kg); AMP-treated with ampicillin (100mg/Kg); AMC group-treated with amoxicillin + clavulanic acid (100mg/kg); CLI-treated with clindamycin (60mg/kg). After euthanasia, the jaws were collected and processed for (1) histological and histometric analysis using hematoxylin and eosin staining, (2) analysis of collagen fibers using Picrosirius Red staining and (3) bacteriological analysis using Brown-Brenn staining. The data were analyzed statistically (p < 0.05). AP induction was confirmed in all groups. The AMC group had the lower intensity of inflammatory infiltrate (p = 0.028) and less periapical bone resorption compared to control (p = 0.006). Regarding collagen maturation, PSR staining revealed a predominance of mature collagen fibers in all groups. The AC and AMC groups had the lower amount of mature fibers and the highest amount of immature fibers, compared to all other groups (p < 0.001). All groups showed bacterial contamination; however, the AC and AMC groups showed a lower extent of bacterial contamination compared to the control (p < 0.001). It can be concluded that systemic antibiotic therapy influences the development and progression of induced AP.

Chenopodium Ambrosioides Linn Mitigates Bone Loss in Rats with Periodontitis.

Periodontitis is an inflammatory disease that causes bone loss. Some patients do not respond well to the classic treatment and need therapies that minimize bone loss, the main sequel of the disease. Chenopodium ambrosioides L. has stood out due to its anti-inflammatory and anti-oxidative activities. However, no study has yet investigated its effect on periodontitis. This study aimed to evaluate the bone protective effect of Chenopodium ambrosioides L. (CAL) extract on ligature-induced periodontitis model in rats. For this, a pre-clinical assay was performed, using male Wistar rats divided into 3 groups: Naive (N) (n=6), not submitted to any procedure; Saline (SAL) (n=6), submitted to ligature-induced periodontitis and receiving 2 ml/kg of 0.9% saline solution; and CAL extract, which was subdivided into 3 subgroups (n=6/subgroup) receiving the CAL at 3 (CAL3), 10 (CAL10) or 30 mg/kg (CAL30). All agents were given, by oral gavage, 30 min before periodontitis induction and daily until euthanasia (11th day). By then, maxillae were removed for macroscopic, histological, and histometric analyses. Kidneys, liver, and stomach were collected to evaluate the safety of CAL extract. High-performance liquid chromatography (HPLC) assay was used to investigate the flavonoid content in the extract. Chenopodium ambrosioides L. extract at 30mg/kg showed a reduction by 58% in bone loss marked by an increase (+35%) in the number of osteoblasts and a reduction (-51%) on the number of osteoclasts (p< 0.05). No significant alteration in the liver, kidney, or stomach was seen. Rutin was the main flavonoid found. In summary, it was observed that Chenopodium ambrosioides L. extract has shown important anti-inflammatory and bone anabolic and anti-resorptive properties without causing toxicity in the main organs. Rutin, as the main flavonoid of the extract, seems to be responsible for the beneficial effect of this agent.

Evaluation of Porcine Collagen Membranes Used with Guided Bone Regeneration for Critical Defects: A Histological, Histomorphometric, Immunohistochemical, and Inflammatory Profile Analysis.

The objective of this study was to compare the effectiveness of two porcine collagen membranes of different origin used for guided bone regeneration procedures. Resorbable collagen membrane from porcine dermis (Bio-Gide, Geistlich Pharma AG, Wolhusen, Switzerland) and resorbable collagen membrane from porcine pericardium (Jason, Institut Straumann AG, Peter Merian-Weg, Switzerland) were evaluated; histological, histometric, immunohistochemical, and inflammatory profile analyses were performed. The study was carried out on critical defects created in the calvaria of 72 rats (Rattus norvegicus albinus, Wistar variety) divided into three groups: coagulum group (Co), porcine pericardium group (JS), and porcine collagen group (BG). The defects were filled with clot, over which the membranes were placed. The animals were euthanized 7, 15, 30, and 60 days after surgery. The Shapiro-Wilk test was used to assess data distribution. Analysis of variance (ANOVA) and the Bonferroni multiple comparison test were used to compare the differences across the mean values of the variables. Nonparametric tests, Mann-Whitney and Wilcoxon W, were used for the quantitative analysis of the inflammatory profile. A significance level of 5% (p < 0.05) was adopted with a confidence interval of 95%. SPSS software version 2.0 was used. A total of 1,008 analyses were performed on 288 histological slides. It was noted that both types of collagen membranes used in this study were effective for the guided bone regeneration procedure, with a greater proportion and thickness of bone formation among recipients of the BG (735 points, p = 0.021). This membrane also had greater permeability (62.25). The animals in the JS group, which received the porcine pericardial membrane, showed early and accelerated bone formation from early bone tissue, milder osteopontin and osteocalcin levels, and greater inflammatory reaction (86.4). The collagen membrane from porcine dermis demonstrated a more orderly and physiological repair process, while the porcine pericardial membrane presented a more accelerated repair process that did not remain constant over time.

New Bone Growth into Surgical Bone Defects Grafted with a Novel Cultivated Coral Graft: A Histomorphometric Study in Rabbit Calvarias.

To evaluate a new cultivated coral graft (CCG) in an in vivo experimental guided bone regeneration (GBR) procedure. The calvarias of eight rabbits were surgically exposed, and circular defects 8 mm in diameter were prepared. One defect was filled with CCG particles (experimental group); the contralateral defect (control group) was spontaneously filled by blood clot. All defects were covered with a collagen membrane. Subjects were euthanized after 8 weeks. Histologic observations of the defects showed similar bone growth patterns in both experimental and control osteotomies. In the experimental defects, no traces of coral particles were observed. Histometric analysis showed denser bone in the pristine zone (65%-66%) than in the peripheral zone for both the control (50%) and experimental defects (31%) (P &#61; not significant). The new bone percentage was reduced from the peripheral zone toward the middle and the center of the defect (31%, 32%, and 27%, respectively) as the distance from the peripheral pristine bone borders increased. The existing data supports the complete degradation of CCG as a spacemaintaining scaffold for GBR procedures.

Temporal and spatial variability of phloem structure in Picea abies and Fagus sylvatica and its link to climate.

Using a unique 8-year data set (2010-2017) of phloem data, we studied the effect of temperature and precipitation on the phloem anatomy (conduit area, widths of ring, early and late phloem) and xylem-ring width in two coexisting temperate tree species, Picea abies and Fagus sylvatica, from three contrasting European temperate forest sites. Histometric analyses were performed on microcores taken from tree stems in autumn. We found high interannual variability and sensitivity of phloem anatomy and xylem-ring widths to precipitation and temperature; however, the responses were species- and site-specific. The contrasting response of xylem and phloem-ring widths of the same tree species to weather conditions was found at the two Slovenian sites generally well supplied with precipitation, while at the driest Czech site, the influence of weather factors on xylem and phloem ring widths was synchronised. Since widths of mean annual xylem and phloem increments were narrowest at the Czech site, this site is suggested to be most restrictive for the radial growth of both species. By influencing the seasonal patterns of xylem and phloem development, water availability appears to be the most important determinant of tissue- and species-specific responses to local weather conditions.

Ozone therapy improves early stages of osseointegration in ovariectomized rats.

the aim of this study was to analyze the influence of ozone therapy (OZN) on peri-implant bone repair in critical bones by installing osseointegrated implants in the tibia of ovariectomized rats. ovariectomy was performed on 30 Wistar rats, aged six months (Rattus novergicus), and, after 90 days, osseointegrated implants were installed in each tibial metaphysis. The study groups were divided into the animals that received intraperitoneal ozone at a concentration of 700 mcg/kg - OZ Group (n=15) - and a control group that received an intraperitoneal saline solution and, for this reason, was named the SAL group (n=15). The applications for both groups occurred during the immediate post-operative period on the 2nd, 4th, 6th, 8th, 10th, and 12th day post-surgery. At various stages (14, 42, and 60 days), the animals were euthanized, and tests were performed on their tibiae. These tests include histomorphometric and immunohistochemical analyses, computerized microtomography, sampling in light-cured resin for calcified sections, and confocal microscopy. The obtained data were then analyzed using One-way ANOVA and the Shapiro-Wilk, Kruskal-Wallis, and student t-tests (P<0.05). our findings indicate that the OZ group (3.26±0.20 mm) showed better cellular organization and bone neoformation at 14 days (SAL group, 0.90±1.42 mm) (P=0.001). Immunohistochemistry revealed that osteocalcin labeling was moderate in the OZ group and mild in the SAL group at 14 and 42 days post-surgery. The data from the analysis of calcified tissues (microtomography, histometric, and bone dynamism analysis) at 60 days showed no statistically significant differences between the groups (P=0.32). it was concluded that ozone therapy anticipated the initial phases of the peri-implant bone repair process.

Unveiling the Therapeutic Potential of Systemic Ozone on Skin Wound Repair: Clinical, Histological, and Immunohistochemical Study in Rats.

This study sought to examine the effects of systemic ozone (O3) treatment on the healing of skin wounds induced on the dorsal surface of Wistar rats. The skin wounds were created using a 10 mm round punch following the sagittal medial plane in 72 rats. Then, the animals were randomly assigned to four groups, each receiving the following treatments: group C, which did not undergo treatment with the O3/O2 mixture; group OZ0.3, administered the O3/O2 mixture at a dose of 0.3 mg/kg; group OZ0.7, given the O3/O2 mixture at a dose of 0.7 mg/kg; and group OZ1.0, provided with the O3/O2 mixture at a dose of 1.0 mg/kg. Six animals from each group were euthanized at 7, 14, and 21 days postoperatively. Clinical, histological, histometric, and immunohistochemical (IHC) analyses were accomplished. Data from clinical and histometric assessments revealed that OZ0.7 and OZ1.0 demonstrated more favorable healing, with greater wound contraction observed in the OZ1.0 group at 14 and 21 days. Histologically, the OZ1.0 group exhibited aspects consistent with an accelerated tissue repair process. IHC analysis revealed greater vascular endothelial growth factor (VEGF) immunostaining in the OZ0.7 (7 days) and OZ1.0 (7 and 14 days) groups compared to the C group. Expression of transforming growth factor beta-1 was significantly increased in the OZ0.7 (14 days) and OZ1.0 (7 and 14 days) groups compared to the C group. In conclusion, our data suggest that systemic use of O3 enhanced tissue repair in cutaneous wounds in a dose-dependent manner, with concentrations of 1.0 mg/kg providing the most beneficial effects. Furthermore, the results of this study implicate the use of O3 for the treatment of skin wounds aiming at improving the healing process over time. Our findings suggest the use of O3 as a viable alternative to enhance wound healing and repair.

Effect of local and systemic administration of atorvastatin for improving bone healing on critical defects.

This study aimed to evaluate the impact of atorvastatin, administered both locally and systemically, on critical defects in the calvaria of rats. Thirty-six adult rats were randomly assigned to three groups, with all bone defects covered by a collagen membrane. The groups received different treatments: distilled water (GAD), where membranes were soaked in distilled water; systemic application of atorvastatin (GAS) at a dosage of 3.6mg/kg/day through gavage; and local application of atorvastatin (GAL). After 14 and 28 days, all animals were euthanized, and various assessments were conducted, including histometric analysis, measurement of linear residual defect, evaluation of newly formed bone area, determination of membrane and soft tissue area, cell count, and immunohistochemical analysis. Group GAS exhibited a significant reduction in residual defect compared to the other groups (p<0.05) and a lower number of osteocytes (p<0.05) in comparison with other groups. On day 28, both GAL and GAS groups showed a higher number of inflammatory cells compared to GAD (p<0.05). Immunolabeling of CD31 was similar for both groups, but in the case of osteocalcin, there was a significant increase in labeling for groups GAS and GAL between days 14 and 28 postoperative (p<0.05). In conclusion, systemic atorvastatin demonstrated enhanced osteogenesis in critical calvaria defects in rats, suggesting its efficacy in promoting bone regeneration without exerting a notable anti-inflammatory effect.

Morphohistometric and steroidogenic parameters during testicular and epididymal differentiation in cavy (Galea spixii) fetuses

Sexual differentiation and steroidogenic mechanisms have an important impact on postnatal gonadal phenotypic development. Thus, establishing the activities that lead to male phenotypic development can provide a better understanding of this process. This study examined the prenatal development of cavies to establish morphological and histometric development patterns and protein and enzyme immunolocalization processes that are responsible for androgen synthesis in the testes and epididymis. Histological and histometric analyses of the diameter of the seminiferous cords and epididymal ducts of male fetuses on Days 25, 30, 40, and 50 were performed, as well as immunohistochemistry of the steroidogenic enzymes 5α-reductase and 17β-HSD, the androgen receptor, and the anti-Müllerian hormone (AMH). Our findings showed a cellular grouping of gonocytes from Day 30 onward that was characteristic of the seminiferous cord, which was not present in the lumen at any of the studied dates. From Day 50 onward, the differentiation of the three anatomical regions of the epididymis was evident, the head (caput), body (corpus), and tail (cauda), with tissue distinctions. Furthermore, the diameters of the seminiferous cords and epididymal ducts significantly increased with age. On Day 50, the tail showed the greatest diameter of the three regions. The Sertoli and Leydig cells exhibited AMH immunoreactivity at all dates. In addition, the Leydig cells and epididymal epithelial tissue were immunopositive for 5α-reductase, 17β-HSD, and the androgen receptor; therefore, these factors influenced the development and maintenance of the testis and epididymis during cavy prenatal development.

Enhanced periodontal tissue healing via vascular endothelial growth factor expression following low-level erbium-doped: yttrium, aluminum, and garnet laser irradiation: In vitro and in vivo studies.

This study aimed to investigate the effects of low-level erbium-doped: yttrium, aluminum, and garnet (Er:YAG) laser irradiation on periodontal tissue healing and regeneration through angiogenesis in vivo and in vitro studies. Intrabony defects were surgically created in the bilateral maxilla molar of rats. The defects were treated by open flap debridement (OFD)with Er:YAG laser, including low-level laser irradiation (LLLI) to bone and blood clot surfaces, or conventional procedures. The mRNA expression of vascular endothelial growth factor (VEGF) in the surgical sites was quantified using real-time polymerase chain reaction. The decalcified specimens were prepared for histometric analysis. Also, LLLI was performed on human umbilical vein endothelial cells to evaluate the effects on angiogenesis. Cell proliferation, VEGF expression, and tube formation were assessed. In addition, capsazepine (CPZ), a selective inhibitor of transient receptor potential vanilloid 1 (TRPV1), treatment was performed before LLLI for the same assays. OFD using Er:YAG laser did not generate thermal damage on bone or root surfaces. LLLI accelerated hemostasis by coagulation of the superficial layers of blood clots in the laser-treated group. Postoperative healing was sound in all animals in both groups. VEGF expression and bone formation were significantly increased in the laser-treated group compared to those in the conventional treatment group. In vitro, cell proliferation and VEGF expression were significantly increased in the LLLI group compared to the control group. Tube-formation assays showed that LLLI significantly promoted angiogenesis. CPZ treatment significantly suppressed VEGF expression and tube formation following LLLI. This study suggests that Er:YAG laser irradiation may promote periodontal tissue healing by enhancing angiogenetic effect of endothelial cells via TRPV1.