Search Results (499)

Apatite pastes derived from eggshell waste (BAp) were implanted onto the calvarial bone of rats, and bone formation was evaluated using X-ray μ-computed tomography (CT) and histological evaluation. BAp was mixed with distilled water to prepare a paste. Monoclinic hydroxyapatite of mineral resources (HAp) was used as a control. A 5 mm diameter PTFE (polytetrafluoroethylene) tube was filled with apatite pastes and implanted in the calvarial bone of 9-week-old Sprague Dawley rats for 8 weeks. A larger radiopaque area, similar to that of native bone, was observed in the BAp paste-implanted specimens than that of HAp paste. The bone mineral density (BMD) value of the BAp paste was significantly higher than that of the HAp paste (p < 0.05). In the histological evaluation, new bone formation was noticed from the calvarial side for both apatite specimens, and HAp remained in the PTFE unlike BAp. The bone mass (BM) value of the BAp paste was significantly higher than that of the HAp paste (p < 0.05). SEM and XRD analyses revealed that BAp was microcrystalline and poorly crystalline. The promotion of new bone formation may contribute to the crystallinity and Mg content of BAp. BAp was found to be useful as a bone regeneration material. Full article

Background/Objectives: Guided bone regeneration (GBR) is a regenerative technique used to treat maxillary osseous defects to enable implant placement for prosthetic rehabilitation. It is generally performed with the use of barrier membranes and bone substitute materials of human or animal origin. Here, we report the clinical and histological outcomes of a horizontal GBR, treated using only synthetic biomaterials. Methods: A graft of nanocrystalline hydroxyapatite (NH) embedded in a silica gel matrix was used to fill a horizontal bone defect. The graft was covered with a titanium-reinforced dense polytetrafluoroethylene (TR-dPTFE) membrane, and primary closure was completed and maintained for 10 months. Then, the site was re-opened for membrane removal and implant insertion. During implant bed preparation, a bone biopsy was obtained for histological evaluation. A metal–ceramic crown was fitted, and the 5-year follow-up after prosthetic loading showed clinical and radiographically healthy tissues. Results: Histological examination revealed good integration of the biomaterial into the surrounding tissues, which were composed of lamellar bone trabeculae and connective tissue. New bone formation occurred not only around the NH granules but even inside the porous amorphous particles. Conclusions: The combination of NH and the TR-dPTFE membrane produced good clinical and histological results, which remained stable for 5 years. Full article

Background: Hypothermia has been previously reported to ameliorate acute renal injury induced by ischemia–reperfusion injury (IRI). However, its protective effects against subsequent renal fibrosis remain unclear. Objectives: The aim of this study was to determine whether hypothermia provides protection against renal ischemia–reperfusion injury (IRI), and to elucidate the molecular mechanisms involved. Methods: We used a model of renal fibrosis after ischemia–reperfusion injury in mice. C57BL/6 mice were divided into the following groups: control mice and ischemia–reperfusion injury mice (at 37 °C and at 32 °C). Their kidneys were harvested on day 1, day 3, and day 7 after IRI. The molecular mechanisms were evaluated. Results: The blood urea nitrogen (BUN) levels, serum creatinine (s-Cr) levels, and the histologic renal injury scores were significantly lower in the 32 °C IRI group than in the 37 °C ischemia–reperfusion injury group. In the hypothermic IR group, TGF-β and α-SMA were significantly decreased, while the PGC-1α level was significantly increased. Cold preparation increased the PGC-1α levels in HK2 cells. In TGF-β-treated HK2 cells, cold preparation decreased α-SMA and collagen IV levels. In addition, siPGC-1α in HK2 cells increased α-SMA and collagen IV, despite cold preparation. Conclusions: Hypothermia attenuates renal function deterioration and renal fibrosis in renal IRI mice kidneys. PGC-1α may play a role in hypothermic protection in renal fibrosis after IRI. Full article

Background: To reconstruct and maintain hard tissues over time, it is necessary to follow effective protocols and use appropriate materials. The selection of the graft material and its properties can also affect the final outcomes. For this purpose, numerous graft materials have been suggested. Among the valuable alternatives to these biomaterials, interest in using teeth as graft material has grown in recent years. Aim: The aim of the study was to investigate the histomorphometric outcomes of using tooth-derived materials when used as a bone substitute. Methods: We evaluated the histological results of autologous demineralized tooth graft material prepared using a Tooth Transformer device. A total of 187 histological samples from 186 subjects (52% male and 48% female, with an average age of 56.30 ± 12.97 years) were analyzed. The analysis focused on the total bone volume (BV%), residual tooth material (residual graft, TT%), and vital bone (VB%). The differences between the presence and absence of the resorbable membrane were also analyzed. Results: The amount of VB was 36.28 ± 19.09%, the residual graft TT was 9.6 ± 10.76%, and 46.96 ± 13.85% was the total bone volume (BV). The presence of membrane increased the amount of BV% and reduced the time to produce bone. Conclusions: The procedure using demineralized autologous tooth-derived biomaterial may be a predictable method for producing new vital bone capable of supporting dental implant rehabilitation and the use of membrane allow better results. Full article

Background/Objectives: Indigo naturalis (IN) is a traditional Chinese medicine concocted from medicinal plants such as Baphicacanthus cusia (Nees) Bremek. IN has multifaceted pharmacological activities. Recent research highlights the remarkable efficacy of IN in treating ulcerative colitis (UC). This study investigates the efficacy of Indigo Naturalis prepared using a novel method (NIN) in ameliorating UC. Methods: We have developed a new IN processing technology without the use of lime. Correspondingly, the content of active ingredients has relatively increased in NIN. In this study, dextran sulfate sodium salt (DSS) induced UC models among male KM mice, and the protective effects of NIN on UC were verified. Results: NIN could significantly improve weight loss, diarrhea and prolapse, bloody stools, elevated Disease Activity Index (DAI) and alleviate the colitis symptoms of mice; it could also improve the shortening of colon, disappearance of intestinal crypts, epithelial cell destruction and inflammatory infiltration caused by UC; and it could also significantly reduce the Histological Index (HI). In addition, NIN relieved the inflammatory response by decreasing the content of pro-inflammatory cytokines TNF-α and IL-1β and elevating the content of anti-inflammatory cytokines IL-10 and IL-22. It also restored the intestinal mucosal barrier by increasing the level of MUC2 protein expression at the site of colonic injury. Conclusions: The significant effects of NIN on UC were verified for the first time, suggesting that NIN was worth further developing into a novel therapeutic drug and, necessarily, further safety evaluations and comparisons with traditional IN will help in the application of NIN. Full article

Background/Objectives: Wilms tumors, the most common pediatric renal tumors, are heterogeneous and consist of varying amounts of three components: blastema, epithelium, and stroma. Postoperative chemotherapy is tailored based on risk group classification and stage. Due to this heterogeneity, pathologists perform extensive tumor sampling to ensure accurate classification. Higher-harmonic-generation microscopy (HHGM) is an innovative imaging technique that enables rapid visualization of fresh tissue without preparation or staining. This makes it particularly valuable for sample selection, as the tissue can be reused for further analysis. This study aims to evaluate the accuracy of pathologists in distinguishing normal renal tissue, abnormal renal tissue, and three types of pediatric renal tumors, Wilms tumor, renal cell carcinoma, and congenital mesoblastic nephroma, in HHGM images. Methods: Twenty-nine samples from eighteen patients with a pediatric renal tumor were imaged using an HHG microscope and subsequently processed for histological analysis. Overview images of the samples were acquired at a rate of 10 s per mm², while high-quality images took 1 min per mm². A multi-observer study involving ten international expert pathologists of the SIOP-RTSG was conducted. Results: Pathologists were able to differentiate between normal and abnormal tissue with 100% (29/29) accuracy and correctly identified tumor versus non-tumor tissue with 97% (28/29) accuracy. Conclusions: These results show that HHGM is a highly promising technique for the rapid assessment of pediatric renal tumor samples, particularly for evaluating sample representativeness. Full article

Archived, frozen tongues of 56 bobcats (Lynx rufus) from Oklahoma, USA, were tested for parasites. Testing for Sarcocystis infections included microscopic examination of unstained muscle squashes, histological sections of paraffin-embedded tissues, and molecular characterization. By a combination of both compression methods and histology, sarcocysts were found in 28 (50.0%) of 56 bobcats. In compression preparations, the sarcocyst wall appeared thin and protrusions were not clear. Histologically, tissues were degraded and, in most tongues, only a few sarcocysts were found, except bobcat #35. Multilocus genotyping utilizing 18S rRNA, 28S rRNA, and cox1 genes yielded sequences exhibiting 98–100% identity with both Sarcocystis arctica and Sarcocystis caninum isolates available in the NCBI database. Hepatozoon rufi-like meronts were found in two tongues and is the first report of Hepatozoon infections in bobcats in Oklahoma. Full article

Cellular infiltration into traditional electrospun nanofibers (NFs) is limited due to their dense structures. We were able to obtain polycaprolactone (PCL) NFs with variable and defined pore sizes and thicknesses by using a customized programmed NF collector that controls the moving speed during electrospinning. NFs obtained by this method were tested in vitro and have shown better cell proliferation within the NFs with larger pore sizes. This study investigated in vivo host cell migration and neovascularization within implanted porous PCL NF discs using a mouse pouch model. Four types of PCL NFs were prepared and classified based on the electrospinning speed: NF-zero (static control), NF-low (0.085 mm/min), NF-mid (0.158 mm/min) and NF-high (0.232 mm/min) groups. With the increase in the speed, we observed an increase in the pore area; NF-zero (11.6 ± 6.2 μm²), NF-low (37.4 ± 28.6 μm²), NF-mid (67.6 ± 54.8 μm²), and NF-high (292.3 ± 286.5 μm²) groups. The NFs were implanted into air pouches of BALB/cJ mice. Mice without NFs served as control. Animals were sacrificed at 7 and 28 days after the implantation. Pouch tissues with implanted NFs were collected for histology (n = three per group and time point). The efficiency of the tissue penetration into PCL NF sheets was closely linked to the pore size and area. NFs with the highest pore area had more efficient tissue migration and new blood vessel formation compared to those with a smaller pore area. No newly formed blood vessels were observed in NF-zero sheets up to 28 days. We believe that a porous NF scaffold with a controllable pore size and thickness has great potential for tissue repair/regeneration and for other healthcare applications. Full article

Background/Objectives: Pediatric gonadal tumors are rare tumors, and germ cell tumors (GCTs) are the most common subgroup. GCTs are heterogeneous tumors and have different subtypes that can be either benign or malignant. Therefore, extensive sampling of the resected tumor is required to obtain an accurate diagnosis. Higher harmonic generation microscopy (HHGM) is an innovative imaging technique that enables rapid visualization of fresh tissue without the need for preparation or staining. This makes it particularly valuable for sample selection, as the tissue can be reused for further analysis. This study aims to evaluate the accuracy of pathologists detecting normal gonadal tissue, germ cell tumors, and other pediatric gonadal tumors in HHGM images. Methods: Twenty-eight samples of twenty-two patients with a germ cell tumor or other gonadal tumor were imaged with the HHG microscope and subsequently processed for histology. Overview images of the samples were made in 10 s per mm², and high-quality images in 1 min per mm². A multi-observer study was performed with five expert pathologists. Results: Pathologists were able to differentiate between tumor and non-tumor tissue with an accuracy of 75% (21/28) on the HHGM images versus an accuracy of 89% (25/28) on the corresponding histology. Discrepancies mainly concerned teratoma cases for HHGM as well as H&E, indicating that sampling errors of these heterogeneous tumors affected the outcomes of this study adversely. Conclusions: Although the sample size was limited by the rarity of these tumors, our data show that HHGM is a promising technique for the rapid assessment of pediatric gonadal tumor samples, particularly in evaluating their representativeness. Full article

In adults, scars are formed after deep skin wound injuries like burns. However, the fetal microenvironment allows for scarless skin regeneration. One component that is abundantly present in the fetal extracellular matrix is hyaluronan (HA). To study whether biomaterials with HA improve wound healing, type I collagen scaffolds with and without HA were prepared and characterized. Their immune effect was tested using macrophages and their phenotypes were analyzed through cell surface markers and cytokine expression after 48 h. Since fibroblasts are the main cellular component in the dermis, adult, fetal and eschar-derived cells were cultured on scaffolds for 14 days and evaluated using histology, gene and protein expression analyses. Biochemical assays demonstrated that HA was successfully incorporated and evenly distributed throughout the scaffolds. Macrophages (M0) cultured on Col I+HA scaffolds exhibited a profile resembling the M2c-like phenotype (CD206^high, CD163^high and IL10^high). HA did not significantly affect gene expression in adult and fetal fibroblasts, but significantly reduced scarring-related genes, such as transforming growth factor beta 1 (TGFB1) and type X collagen alpha 1 chain (COL10A1), in myofibroblast-like eschar cells. These findings highlight the potential of incorporating HA into collagen-based skin substitutes to improve the wound healing response. Full article

Background: Scald burns pose significant morbidity, and effective topical treatments remain a clinical priority. Burn injuries pose a significant clinical challenge due to the prolonged inflammation and high infection risk. Traditional treatments focus on moisture retention and infection prevention, but biocompatible formulations such as hydrogels and oleogels offer advantages. Hydrogels hydrate, cool, and promote epidermal regeneration, while oleogels form a lipid barrier that enhances the absorption of lipophilic bioactive compounds. There is an increasing demand for novel topical alternatives that can effectively improve wound healing by modulating the inflammatory cascade, accelerating epithelial and dermal regeneration, and restoring barrier function. Objective: This study aimed to determine the most effective plant-based topical formulations for enhancing second-degree scald burn wound healing. Methods: Utilizing a standardized rat model, we compared 21 distinct topical formulations, consisting of oleogel and hydrogel bases enriched with extracts from Boswellia serrata (frankincense), Ocimum basilicum (basil), Sambucus nigra flower (elderflower), and Galium verum (lady’s bedstraw). Second-degree burns were uniformly induced in 24 Wistar rats using boiling water (100 °C for 8 s) using the RAPID-3D device, a validated 3D-printed tool that ensures reproducible burns through controlled exposure to boiling water. Post-burn, rats were divided into three equal subgroups, and topical formulations were applied daily. Wound healing efficacy was evaluated through wound surface area measurements, transepidermal water loss (TEWL), skin hydration, sebum production, pigmentation, inflammation (erythema), skin perfusion, and histological parameters at multiple timepoints (days 1, 4, 9, 14, and 21 post-burn induction). Results: Statistical analyses indicated significant advantages of oleogel-based formulations over hydrogel-based formulations. Specifically, formulations containing Boswellia serrata and Ocimum basilicum extracts significantly reduced wound size and inflammation, improved skin hydration, and decreased melanin production by days 9 and 21 (p < 0.05). Conclusions: These findings underscore the potential clinical value of oleogel-based topical preparations containing specific plant extracts for improving scald burn wound healing outcomes, warranting further clinical evaluation. Full article

Dendritic cells (DCs) are major contributors to generating an effective immune response due to their ability to present antigens to T cells. Recently, nanoparticles have been widely used in different medical applications, such as drug-delivery systems, to enhance the function of impaired immune cells. Objectives: This research aims to develop an effective antitumor DC-based vaccine by adsorption of chitosan-nanoparticles (CH-NPs) onto DCs. Methods: Undifferentiated mouse bone marrow progenitor cells were differentiated into mature DCs using cytokines and lipopolysaccharides. CH-NPs were prepared using the ionic gelation method and subsequently used to coat the stimulated DCs. The MTT assay was employed to assess the cytotoxicity of all formulations. To compare the antitumor effect of CH-NPs, DCs, and DCs-CH-NPs, mice were divided into five groups and injected with the respective vaccine formulations. Following immunization, flow cytometry was used to analyze DC and CD4⁺ T cell activation in blood and spleen tissues. Histological samples from the spleen and lymph nodes were also collected. Results: Our findings show that co-stimulatory molecules CD80/CD86 and the DC maturation marker CD83 were upregulated in the vaccinated DCs, indicating their maturation. Moreover, CD83, CD11c, and MHC-II were upregulated in blood and spleen samples in vivo. The DC-CH-NPs vaccinated group had a higher mean percentage of CD83 expression in blood samples (76.7 ± 17.1) compared to the DCs group (47.7 ± 11.0) and the CH-NPs group (37.7 ± 8.6). DC markers, particularly CD83, were highly expressed in spleen samples. Additionally, the DC-CH-NPs vaccinated group had a significantly higher number of CD4⁺ T cells (MFI = 26.1 ± 2.3) compared to the DCs (18.6 ± 1.6) and CH-NPs (13.3 ± 1.4) groups. Conclusions: The present study concludes that the DC-CH-NPs vaccine formulation can induce a potent in vivo immune response. These data may provide valuable insights for developing effective delivery systems for antitumor vaccines. Full article

Manual processing of lipoaspirate can enhance stem cell concentration, thereby improving the take rate, which still represents a major challenge in autologous fat transfer. However, since the preparation consists of many manual steps that are difficult to standardize, we investigated the influence of residual tumescent solution on the macroscopic and microscopic outcome of the mechanically processed lipoaspirate. Additionally, we investigated whether sedimentation followed by vacuum filtration of the aqueous phase could accelerate processing by replacing the initial centrifugation step. Samples with more than 5% remaining aqueous phase show no clearly defined oil phase, preventing any volume reduction. In contrast, all centrifuged samples produced a clear oil phase. The remaining tissue, as confirmed by both histology and viability assays, was superior to nanofat. Although sedimentation and filtration in the LipoCollector did not sufficiently separate enough aqueous phase from the lipoaspirate, tissue viability was significantly higher compared to our control container. Our findings indicate that centrifugation remains essential for effective aqueous phase separation and further mechanical processing, while the automatic filtration may enhance processing efficiency. These results indicate that further work is needed to simplify mechanical processing, as the outcome can be significantly influenced by parameters such as tumescent impurities. Full article

This study aimed to investigate the effect of Artemisia absinthium extract on testicular dysfunction in rats and explain the involvement of the androgen receptor signaling pathway as a biomarker in maintaining fertility during sodium fluoride (NaF) treatment. Thirty-two male rats were divided equally into four groups and received treatment for 60 days. The control group (I) received normal saline; group II received Artemisia extract at 100 mg/kg b.w.; group III received NaF at 12 mg/kg b.w. orally; and group IV received NaF (12 mg/kg b.w.) and Artemisia extract (100 mg/kg b.w.). The testis weights and the lipid peroxidation, luteinizing hormone (LH), follicle-stimulating hormone (FSH), and testosterone levels were estimated. The genital organs were prepared and immunoreacted with a receptor. Histomorphometric analyses were performed to obtain the diameter of the seminiferous tubules and the height of the germinal epithelia in the testes. The results showed that exposure to NaF caused a significant increase in testis weight and malondialdehyde (MDA) and a decrease in serum LH, FSH, and testosterone concentrations compared to the control group, while extract administration induced an increase in the levels of these hormones in group IV. Testicular histological and histomorphometric changes were observed in group III: degenerative seminiferous tubules with vascular congestion, disorganization of the germinal layer, and decreased seminiferous tubule diameter and germinal epithelium height. The expression of androgen receptors in the testes of the NaF-treated rats was significantly reduced. In contrast, these testicular histological changes were ameliorated in rats treated with the extract. The results allow us to conclude that the administration of Artemisia absinthium confers positive effects on male reproductive function by inhibiting fluoride, maybe via ameliorative testicular function. Full article

Visualizing the microscopic structure of plants in vivo, non-invasively, and in real-time is the Holy Grail of botany. Optical coherence tomography (OCT) has all the characteristics necessary to achieve this feat. Indeed, OCT provides volumetric images of the internal structure of plants without the need for histological preparation. With its micrometric resolution, OCT is commonly used in medicine, primarily in ophthalmology. But it is seldom used in the field of botany. The aim of the present work is thus to review the latest technical development in the field of OCT and to highlight its current use in botany, in order to promote the technique and further advance research in the field of botany. Full article