Histomorphological Evaluation Research Articles

Histopathological and Immunohistochemical Evaluation of Bone Marrow Biopsy in Myelodysplastic Syndromes

SUMMARY In myelodysplastic syndromes (MDS), bone marrow biopsy (BMB) is now considered part of a multidisciplinary approach to bone marrow (BM) examination that must include cytological and cytogenetic assessment, while flow-cytometry and molecular studies can provide additional useful information. Undervalued in the past, BM histology has recently grown in importance in terms of diagnosis (i.e., MDS with BM fibrosis, hypoplastic MDS) and prognostic assessment (i.e., clusters of CD34+blasts). Thus, BM histology must be considered complementary and not overlapping with the results of the other methods of BM study. Moreover, in recent years, numerous clinico-pathological studies have evidentiated, codified and standardized a series of histopathological parameters that are now considered essential for an effective and clinically useful histomorphological evaluation of the BMB. The aim of this work is to describe the histopathological and immunohistochemical approach to BMB that the pathologists of the Lombard Hematological Network (Rete Ematologica Lombarda [REL]; Italy) are carrying out on a standardized and shared basis in MDS patients.

CDKN2A and MC1R variants influence dermoscopic and confocal features of benign melanocytic lesions in multiple melanoma patients

Non-invasive diagnostic tools are effective in the histomorphological study of melanocytic lesions. The role of melanoma susceptibility genes on melanocytic nevi histopathological features is not clear. The current study aimed to correlate genetic alterations and histomorphological features of melanocytic nevi. Clinical, dermoscopic and confocal features of 34 multiple melanoma patients and 34 controls were compared. Among patients with melanoma, carriers of CDKN2A mutations and/or MC1R variants, and wild-type genes were also compared. In patients with melanoma, a lighter phototype (P=0.051), a higher number of nevi (P<0.01) and clinically atypical nevi (P<0.01) were observed. At dermoscopy, these nevi showed a complex pattern (P=0.011), atypical network (P=0.018) and irregular pigmentation (P=0.037); at confocal, an irregular meshwork pattern (P=0.026) with atypical nests (P=0.016) and an inflammatory infiltrate (P=0.048) were observed. Among patients with melanoma genetically tested, CDKN2A G101W mutation carriers were more frequently younger (P=0.023), with clinically atypical nevi (P=0.050), with cytological atypia (P=0.033) at confocal. G101W mutation and MC1R variants carriers showed hypopigmented nevi (P=0.002) and, at confocal, roundish cells infiltrating the junction (P=0.019). These data suggest an influence of CDKN2A mutation and MC1R variants in the development of dysplastic melanocytic lesions. Non-invasive histomorphological evaluation, together with genetic studies, improves melanoma risk identification and early diagnosis, for a patient-tailored management.

Bone marrow findings in HIV-positive patients with Kaposi sarcoma herpesvirus-associated multicentric Castleman disease.

Kaposi sarcoma herpesvirus (KSHV), also known as human herpesvirus-8, is associated with 1 form of multicentric Castleman disease (MCD) and is the etiologic agent for most MCD in human immunodeficiency virus (HIV)-infected patients. Diagnosis is usually determined by lymph node biopsy. Bone marrow findings in KSHV-MCD are not well characterized. We conducted histomorphologic and immunohistochemical evaluation of bone marrow biopsy specimens in HIV-infected patients with KSHV-MCD, including evaluation for KSHV latency-associated nuclear antigen. Findings were correlated with clinical features and KSHV viral load. Reactive plasmacytosis was the predominant feature. Lymphoid aggregates were less common and not diagnostic of KSHV-MCD. Forty-eight percent of cases contained scattered KSHV-infected mononuclear cells. Although patients were generally cytopenic, bone marrow biopsy specimens were normocellular to hypercellular except in patients receiving hematotoxic therapy. Bone marrow biopsy specimens in KSHV-MCD patients recapitulate findings of interleukin-6 excess. In patients with HIV, unexplained cytopenias, and bone marrow plasmacytosis, evaluation for KSHV-MCD is warranted.

Alterations in digestive enzyme activities during the development of diet-induced enteritis in Atlantic salmon, Salmo salar L.

This study followed the time-related changes in intestinal digestive physiology of post-smolt Atlantic salmon (Salmo salar L.; initial weight 500–600g) during the development of soybean meal induced enteritis. Intestinal tissue and content (chyme) were sampled after 1, 2, 3, 5, 7, 10, 14, 17, and 21days of dietary exposure to a 20% extracted soybean meal-containing (SBM) diet. Fish fed a fishmeal-based control diet sampled on day 0 (prior to SBM feeding) served as controls. From the different intestinal regions, histomorphological evaluation was conducted, activities of the pancreatic enzymes trypsin, chymotrypsin, elastase and lipase, as well as leucine aminopeptidase (LAP) were determined in chyme, and the brush border membrane (BBM) enzymes LAP and maltase were measured in homogenates of the intestinal wall. Epithelial cell functional differentiation was visually assessed using immunohistochemical expression of carbonic anhydrase 12 (CA-12). Histological signs of SBM-induced inflammation were observed in the distal intestine (DI) from day 5. At each given sampling time, pancreatic enzyme activities generally decreased aborally along the intestinal tract. Within each intestinal region, pancreatic protease activities tended to increase with exposure time to the SBM diet, with the greatest activity increases observed in the DI. The magnitude of the decrease in activity between the most proximal region and the most distal decreased with SBM exposure. DI tissue weight and BBM maltase and LAP activities were significantly reduced from day 1–2 and continued to decrease progressively with increasing SBM exposure time. Immunohistochemical staining of CA-12 also diminished over time starting at day 5. By day 7, very little CA-12 staining was observed. The data suggests an immediate and generalized physiological response to a change in diet and/or feed intake in all regions of the intestine, as well as a more localized functional loss in the DI that progressed with the development of SBM-induced inflammation from day 5. The diet change appeared to cause a rapid pancreatic response with increased secretion of all the enzymes investigated, while the longer term responses of the pancreatic proteases differed from that of lipase.

Histomorphological and immunofluorescence evaluation of clear corneal incisions after microcoaxial phacoemulsification with 2.2 mm and 1.8 mm systems

To compare changes in the incision's histomorphology and denaturation of collagen I in rabbit eyes having microcoaxial phacoemulsification through 2.2 mm and 1.8 mm incision-compatible systems. Randomized experimental trial. Iladevi Cataract & IOL Research Centre, Ahmedabad, India. Thirty rabbit eyes were randomized into Group 1 (microcoaxial phacoemulsification through 2.2 mm incisions using Infiniti system [torsional ultrasound]) and Group 2 (microcoaxial phacoemulsification through 1.8 mm incisions using Stellaris system [longitudinal ultrasound]). Each group was then divided into 3 subgroups of 5 eyes each based on 1 of the 3 intervention options: phacoemulsification only, intraocular lens (IOL) insertion only, and phacoemulsification with IOL insertion. Left eyes were randomized for microcoaxial phacoemulsification, and right eyes were treated as controls. After phacoemulsification, eyes in Group 1 showed loss of epithelium at the roof of the incisions and Descemet membrane detachment at the floor of the incisions. These findings did not change after IOL insertion. After phacoemulsification, eyes in Group 2 showed loss of epithelium, but Descemet membrane remained attached. There was a longitudinal split in the incision's stroma in the direction of internal entry. The stromal damage increased after IOL implantation. Immunofluorescence studies showed no obvious irregularities in the arrangement of collagen I in either group. A dot blot analysis showed significant denaturation of collagen I in Group 2. The histomorphology of the 2.2 mm system incision showed localized Descemet membrane detachment and endothelial cell loss. The 1.8 mm system incision showed exaggerated stromal damage after IOL insertion.

Fourier transform infrared imaging analysis in discrimination studies of bladder cancer

Bladder carcinoma represents more than 4% of all cancer diseases in Austria. The histomorphological evaluation is invasive and remains a subjective and time consuming technique. On account of this it is necessary to find novel non-invasive approaches which support the pathologists for histological recognition to identify malignancy at an early stage. In the present study, Fourier transform infrared (FTIR) microscopic imaging was combined with univariate and multivariate data analysis methods to study bladder carcinoma tissue sections in detail. The possibility to collect IR spectra of bladder carcinoma tissue sections employing an optimized analytical protocol is demonstrated. The correlation between FTIR microscopic imaging and the morphological tissue features obtained by histological staining of the sections demonstrated that many histomorphological tissue patterns can be visualized in the colour images. The routine generation of high quality imaging data is enabled because of the combination of FTIR technology and optimized sample preparation techniques. This opens a new quality of spectroscopic analyses of cancerous tissue, allowing exploration of molecular changes associated with the histopathological morphology.

Phenotypic Modulation of Cell Types around Implanted Polyethylene Terephthalate Fabric in Rabbit Muscle

Whereas the nature of healing reaction in skeletal muscle following implantation of biomaterial has been extensively studied, the extent of variation in cell phenotypes is poorly known. Here, we studied the phenotypic alteration of cell types following injury in skeletal muscle of rabbits implanted with a commonly used biomaterial, polyethylene terephthalate (PET) fabric. Following implantation, histomorphological features were studied after 1, 4, and 12 weeks. Routine objective histomorphological evaluation was supplemented with histochemistry for collagen and immunohistochemistry for proliferating cell nuclear antigen (PCNA), CD34, vimentin, and alpha smooth muscle actin (α-SMA). The extent of reaction was quantified. The foreign body giant cells were found to comprise subpopulations, based on the variation in vimentin detectability or the presence of differentially capable proliferating nuclei (PCNA positive). Many rhabdomyocytes adjacent to the implant were PCNA-positive and some of them showed CD34 positivity. Often, the rhabdomyocytes very near to implanted PET fabric assumed a myofibroblast phenotype as evidenced by vimentin and/or α-SMA positivity at immunohistochemistry. Overall, the results suggested a phenotypic alteration of native cell types following implantation of PET fabric in rabbit skeletal muscle. Quantification of such cell types at the tissue-material interphase in relation to the deposition of collagen may be desirable during safety evaluation of biomaterials by histomorphology.

Ameliorative effect of grape seed proanthocyanidin extract on thioacetamide-induced mouse hepatic fibrosis

The present study was designed to examine the effect of the grape seed proanthocyanidin extract (GSPE) on developing hepatic fibrosis that was induced by thioacetamide (TAA) in mice. Administration of TAA for 9 weeks led to a serious necrosis and apoptosis of the parenchymal cells, which resulted in an accumulation of excessive collagen in the liver and an increase of transformed hepatic stellate cells (HSCs). In addition, the mRNA expression of transforming growth factor β1 (TGF-β1), α-smooth muscle actin (α-SMA), as the marker of the activated HSCs, and α1-(I)-collagen were all up-regulated significantly when compared with the control. However, combined oral administration of GSPE at 100mg/kg suppressed the mRNA expression of TGF-β1 and α-SMA, with decreased collagen accumulation as demonstrated by histomorphological evaluation and quantitative RT-PCR. The mRNA expression of the pro-inflammatory factors, including inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), was remarkably enhanced by TAA treatment. However, their levels displayed a down-regulated trend beyond simultaneous GSPE treatment. Moreover, GSPE administration markedly suppressed lipid peroxidation. In conclusion, as a plant antioxidant, GSPE manifested effective hepatocellular protective action to ameliorate the developing liver fibrosis induced by chronic TAA administration in mice.

Correlation of radio- and histomorphological pattern of pulmonary adenocarcinoma

Recently, a novel classification system based on tumour architecture and with high prognostic impact has been proposed for pulmonary adenocarcinomas (ADCs). For imaging-based prediction of histological ADC subtypes and, thus, prognosis, it is of paramount importance to investigate the correlations of radio- and histomorphological parameters. Associations between histomorphological ADC growth patterns (lepidic, acinar, papillary, micropapillary and solid) and data from pre-operative assessment by computed tomography (CT) imaging of 174 resected pulmonary ADCs were analysed. Margin configuration as well as solidity/ground glass opacity of an ADC was associated with distinct histomorphological ADC growth patterns. Solid-predominant ADCs usually had smooth margins and were also solid in CT scans, while lepidic-predominant ADCs had no predominant margin pattern, were located in the periphery, showed a positive bronchogram and were frequently associated with solidity/ground glass opacity. In addition, nonspherical tumour growth was a negative predictor of overall and disease-specific patient survival. We defined CT morphological parameters that were associated with histomorphological growth patterns of pulmonary ADCs. These data may form the basis for the development of future prognostic algorithms in the palliative setting, which include an integrated evaluation of biopsy histomorphology and CT scan morphology of nonresectable pulmonary ADC.

The protective effects of carnosine in alcohol-induced hepatic injury in rats

Consumption of alcohol leads to oxidative stress in liver by inducing lipid peroxidation. The aim of this study was to investigate the effects of carnosine (CAR) in alcohol-induced liver injury by biochemical and histomorphological evaluations. The rats were divided into four groups, namely, control group, alcohol (AL) group, CAR group and AL+CAR group. Three doses of ethanol (5g/kg, 25% (v/v) in distilled water) were given by nasogastric catheter for twice-a-day. CAR (100mg/kg) was given 1h before the administration of ethanol using the same method. Levels of alanine aminotransferase, aspartate aminotransferase, myeloperoxidase and malondialdehyde were significantly increased in the AL group compared with control, CAR and AL+CAR groups. Glutathione level was significantly decreased in the AL group, while it was increased in the AL+CAR group. Immunoreactivity of caspase-3 and bax increased in the hepatocytes of AL group when compared with control and AL+CAR groups. Expression of bcl-2 was decreased in AL group than AL+CAR group. Under electron microscopy, dense mitochondria, accumulation of lipid, sinusoidal dilatation, vacuolization and decrease in the number of microvilli were observed in AL group, while these findings were markedly less in the AL+CAR group. In conclusion, pretreatment of CAR is effective for recovering biochemical alterations and morphologic damage in the liver of rats treated with ethanol.

Enhancement of Bone Regeneration Using Osteogenic-Induced Adipose- Derived Stem Cells Combined with Demineralized Bone Matrix in a Rat Critically-Sized Calvarial Defect Model

Human adipose tissue contains pluripotent stem cells that are similar to bone marrow-derived stem cells. The present study examined whether osteogenic induced adipose-derived stem cells (ASCs) could enhance the osteogenic capacity of demineralized bone matrix and accelerate bone formation in a rat critically-sized calvarial defect model. Forty Sprague-Dawley rats were divided randomly into four groups containing 10 rats per each group (Control, 0.05 cc fibrin glue (25 mg/ml) and 0.05 cc thrombin (130 U/ml); DBX, control + 0.2 g DBX�; ASC, DBX + 1 x 105 ASCs/g; iASC, DBX + 1 x 105 osteogenic-induced ASCs/g). After osteogenic differentiation of ASCs, alkaline phosphatase and von Kossa staining were performed each week to determine the extent of differentiation and mineralization. An 8-mm critical size circular defect was made in the calvarial bone of each rat. The specimens were harvested 8 weeks after implantation, and radiographic and histological evaluations were carried out. New bone formation was quantified by radiodensitometric analysis of the calvarial sections. Statistical analysis was accomplished using a Mann-Whitney test and Kruskal-Wallis test at a significance level of P < 0.05. Alkaline phosphatase and von Kossa staining showed that the osteogenic-induced ASCs yielded higher osteogenic differentiation at 3 weeks. The calvarial defect was filled more in the iASC group compared to the other groups, as demonstrated by the gross appearance of the specimen and radiologic evaluation. The mean radiodensity of the control, DBX, ASC, and iASC group was 16.78%, 39.94%, 25.58%, and 51.31%, respectively, and these were significantly different (P=0.034). Histomorphological evaluation confirmed that new bone formation was accelerated and enhanced by the osteogenic-induced ASCs. ASCs produced greater osteogenic differentiation at 3 weeks. Osteogenic regeneration was accelerated and enhanced in vivo with the osteogenic-induced ASCs, compared to undifferentiated ASCs. Osteogenic-induced ASCs are an excellent and promising candidate for regenerative medicine and tissue engineering application.

Histomorphological evaluation of mice testis after co-exposure to lead and cadmium

The present study investigates the effects of co-exposure to lead and cadmium on mice testis using histomorphological approach. Male mice were subcutaneously injected with lead chloride on day 1 and cadmium chloride on day 2 (74 and 1 mg/kg body weight, respectively), and kept for 24 h. Vehicle control group was also considered. Mice were then sacrificed and testis were collected and weighed. Samples were fixed on Bouin's solution and processed for histology. The diameter of seminiferous tubules in both groups was calculated using software based on deformable models (SNAKE). The combined exposure of lead and cadmium induced degenerative changes in testis, namely, wavy contour of seminiferous tulules, germ cell loss, and release of immature cells into the lumen. Atrophy of seminiferous tubules was seen in this group, confirmed by a significant ( P <0.001) decrease in the diameter. Cumulative effects of lead and cadmium may have disrupted the blood-testis barrier, then causing the histopathological lesions within testis.

The effects of exposure to electromagnetic field on rat myocardium

Exposure to electromagnetic fields (EMFs) causes increased adverse effects on biological systems. The aim of this study was to investigate the effects of EMF on heart tissue by biochemical and histomorphological evaluations in EMF-exposed adult rats. In this study, 28 male Wistar rats weighing 200-250g were used. The rats were divided into two groups: sham group (n=14) and EMF group (n=14). Rats in sham group were exposed to same conditions as the EMF group except the exposure to EMF. Rats in EMF group were exposed to a 50-Hz EMF of 3mT for 4h/day and 7days/week for 2months. After 2months of exposure, rats were killed; the hearts were excised and evaluated. Determination of oxidative stress parameters was performed spectrophotometrically. To detect apoptotic cells, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining and caspase-3 immunohistochemistry were performed. In EMF-exposed group, levels of lipid peroxidation significantly increased and activities of superoxide dismutase and glutathione peroxidase decreased compared with sham group. The number of TUNEL-positive cells and caspase-3 immunoreactivity increased in EMF-exposed rats compared with sham. Under electron microscopy, there were mitochondrial degeneration, reduction in myofibrils, dilated sarcoplasmic reticulum and perinuclear vacuolization in EMF-exposed rats. In conclusion, the results show that the exposure to EMF causes oxidative stress, apoptosis and morphologic damage in myocardium of adult rats. The results of our study indicate that EMF-related changes in rat myocardium could be the result of increased oxidative stress. Further studies are needed to demonstrate whether the exposure to EMF can induce adverse effects on myocardium.

Rho‐inhibition by local application of c3‐toxin for enhancement of axonal sprouting in a rat end‐to‐side nerve repair model

The aim of the presented study was to investigate nerve regeneration after application of C3-Toxin, a Rho-GTPase inhibitor and to correlate morphometry, neurophysiology, and function in an end-to-side peroneal/tibial nerve repair model of the rat. Twenty rats with a peroneal to tibial end-to-side neurorrhaphy were divided into two groups: 1) control group, 2) C3 fusion toxin group with intrafascicular application of 1 μg/100 μl C3 fusion toxin. Survival time was 8 weeks. Nerve conduction velocities and motor function were analyzed and histomorphological evaluation consisting of measurement of intraneural collagen level, axon count, total nerve area, myelination index, and N-ratio followed. Evaluation of motor function and nerve conduction did not show any statistical differences. Histological analysis revealed higher axon count, thicker myelin sheaths, and higher myelination index in the C3 fusion toxin group (P < 0.001). Comparison of N-ratio and intraneural collagen level were without statistical significance. The current study shows that application of C3 fusion toxin leads to higher myelination and increases axonal sprouting.

Biomechanical and Bone Histomorphological Evaluation of Two Surfaces on Tapered and Cylindrical Root Form Implants: An Experimental Study in Dogs

The aim of this study was to evaluate the early bone response of tapered and cylindrical root form implants with two different surface treatments in fresh extraction sockets after 4 and 8 weeks. Surface treatments and implant design comprised (n = 9 each): tapered with dual acid-etched surface; tapered with dual acid-etched and sandblasted surface (T DAE SB); cylindrical with dual acid-etched surface (C DAE); and cylindrical with dual acid-etched and sandblasted surface (C DAE SB). Implants were placed in the distal sockets of mandibular premolars (2 P2, 3 P3, 4 P4 ) of six beagle dogs, remaining in vivo for 4 and 8 weeks. After sacrifice, the implants were subjected to torque to the point of interface fracture and subsequently nondecalcified for histomorphological study. Statistical analysis was performed by a General Linear Model (GLM) analysis of variance model with a significance level of 5%. Torque to interface fracture was significantly greater for the C DAE SB group than for the other groups (p < .001). Histomorphological analysis showed woven bone formation around all implant surfaces at 4 weeks and its replacement by lamellar bone at 8 weeks. Study time (4 or 8 weeks) did not affect torque measures. The double acid-etched and sandblasted sample surface increased early bone biomechanical fixation of both cylindrical and tapered root form implants. The cylindrical root form implants showed higher torque to interface fracture values when compared with the tapered root form implants. The C DAE SB surface group showed the highest biomechanical fixation values (p < .001).

Effects of Immediate Orthodontic and Orthopedic Forces on Peri-Miniscrew Bones: Histomorphologic and Histomorphometric Assessment in Dogs

Aim. The aim of this study was histomorphologic and histomorphometric evaluation of peri-screw bone after immediate orthodontic and orthopedic forces and comparing them with a control group. Materials and Methods. 18 dual-top miniscrews were inserted in the premolar region of three Iranian dogs. Screws were divided into three groups: (1) the first group undergoing immediate orthodontic force of 300 cN, (2) the second group undergoing immediate orthopedic force of 600 cN and (3) a control group. Screws were explanted with adequate amount of surrounding bone after three months. Bone-screw contact (BSC), percentage of lamellar bone, and percentage of woven bone were evaluated. Wilcoxon and Man Whitney tests were used to analyze the data using SPSS software ver. 15 (α = 0.05). Results. There was no significant difference among the groups in terms of bone-screw contact (P value = 0.42), percentage of lamellar bone (P value = 0.83), and percentage of woven bone (P value = 0.88). Conclusion. By applying orthodontic and orthopedic forces to mini-screws the quality of surrounding bone and osseointegration will not be affected. Clinical Significance. Application of force to mini-screws is helpful in orthodontic-screw therapies.

Loss of sfrp1 promotes ductal branching in the murine mammary gland

BackgroundSecreted frizzled-related proteins (SFRPs) are a family of proteins that block the Wnt signaling pathway and loss of SFRP1 expression is found in breast cancer along with a multitude of other human cancers. Activated Wnt signaling leads to inappropriate mammary gland development and mammary tumorigenesis in mice. When SFRP1 is knocked down in immortalized non-malignant mammary epithelial cells, the cells exhibit a malignant phenotype which resembles the characteristics observed in metastatic breast cancer stem-like cells. However, the effects of SFRP1 loss on mammary gland development in vivo are yet to be elucidated. The work described here was initiated to investigate the role of SFRP1 in mammary gland development and whether SFRP1−/− mice exhibit changes in mammary gland morphology and cell signaling pathways shown to be associated with SFRP1 loss in vitro.Results10 week old nulliparous SFRP1−/− mammary glands exhibited branching with clear lobulo-alveolar development, which normally only occurs in hormonally stimulated mid-pregnant wt mammary glands. Explant cultures of SFRP1−/− mammary glands display increased levels of a well known Wnt signaling target gene, Axin2. Histomorphologic evaluation of virgin glands revealed that by 10 weeks of age, the duct profile is markedly altered in SFRP1−/− mice showing a significantly higher density of ducts with distinct alveoli present throughout the mammary gland, and with focal ductal epithelial hyperplasia. These findings persist as the mice age and are evident at 23 weeks of age. Changes in gene expression, including c-Myc, TGFβ-2, Wnt4, RANKL, and Rspo2 early in mammary gland development are consistent with the excessive hyper branching phenotype. Finally, we found that loss of SFRP1 significantly increases the number of mammary epithelial cells capable of mammosphere formation.ConclusionsOur study indicates that SFRP1 gene is critical for maintaining proper mammary gland development, and that reduced levels of SFRP1 results in hyperplastic lesions and its loss may be a critical event in cancer initiation.

Fourier transform infrared imaging analysis in discrimination studies of squamous cell carcinoma

Oral squamous cell carcinoma (OSCC) of the oral cavity and oropharynx represents more than 95% of all malignant neoplasms in the oral cavity. Histomorphological evaluation of this cancer type is invasive and remains a time consuming and subjective technique. Therefore, novel approaches for histological recognition are necessary to identify malignancy at an early stage. Fourier transform infrared (FTIR) imaging has become an essential tool for the detection and characterization of the molecular components of biological processes, such as those responsible for the dynamic properties of tumor progression. FTIR imaging is a modern analytical technique enabling molecular imaging of a complex biological sample and is based on the absorption of IR radiation by vibrational transitions in covalent bonds. One major advantage of this technique is the acquisition of local molecular expression profiles, while maintaining the topographic integrity of the tissue and avoiding time-consuming extraction, purification, and separation steps. With this imaging technique, it is possible to obtain unique images of the spatial distribution of proteins, lipids, carbohydrates, cholesterols, nucleic acids, phospholipids, and small molecules with high spatial resolution. Analysis and visualization of FTIR imaging datasets are challenging and the use of chemometric tools is crucial in order to take advantage of the full measurement. Therefore, methodologies for this task based on the novel developed algorithm for multivariate image analysis (MIA) are often necessary. In the present study, FTIR imaging and data analysis methods were combined to optimize the tissue measurement mode after deparaffinization and subsequent data evaluation (univariate analysis and MIAs). We demonstrate that it is possible to collect excellent IR spectra from formalin-fixed paraffin-embedded (FFPE) tissue microarrays (TMAs) of OSCC tissue sections employing an optimised analytical protocol. The correlation of FTIR imaging to the morphological tissue features obtained by histological staining of the sections demonstrated that many histomorphological tissue patterns can be visualized in the colour images. The different algorithms used for MIAs of FTIR imaging data dramatically increased the information content of the IR images from squamous cell tissue sections. These findings indicate that intra-operative and surgical specimens of squamous cell carcinoma tissue can be characterized by FTIR imaging.

Investigating the Effect ofAloe veraGel on the Buccal Permeability of Didanosine

The buccal mucosal route offers several advantages but the delivery of certain drugs can be limited by low membrane permeability. This study investigated the buccal permeability properties of didanosine (ddI) and assessed the potential of Aloe vera gel (AVgel) as a novel buccal permeation enhancer. Permeation studies were performed using Franz diffusion cells, and the drug was quantified by UV spectroscopy. Histomorphological evaluations were undertaken using light and transmission electron microscopy. The permeability of ddI was concentration-dependent, and it did not have any adverse effects on the buccal mucosae. A linear relationship (R² = 0.9557) between the concentrations and flux indicated passive diffusion as the mechanism of drug transport. AVgel at concentrations of 0.25 to 2 %w/v enhanced ddI permeability with enhancement ratios from 5.09 (0.25 %w/v) to 11.78 (2 %w/v) but decreased permeability at 4 and 6 %w/v. Ultrastructural analysis of the buccal mucosae treated with phosphate buffer saline pH 7.4 (PBS), ddI/PBS, and ddI/PBS/AVgel 0.5 %w/v showed cells with normal plasmalemma, well-developed cristae, and nuclei with regular nuclear envelopes. However, cells from 1, 2, and 6 %w/v AVgel-treated mucosae showed irregular nuclear outlines, increased intercellular spacing, and plasmalemma crenulations. This study demonstrates the potential of AVgel as a buccal permeation enhancer for ddI to improve anti-HIV and AIDS therapy.

Interleukin-10 and regeneration in an end-to-side nerve repair model of the rat

End-to-side (ETS) neurorrhaphy is an option in peripheral nerve surgery. The aim of this study was to investigate whether the application of the anti-inflammatory cytokine interleukin-10 (IL-10) reduces scarring and thus enhances nerve regeneration in an ETS peroneal/tibial nerve lesion model of the rat. Twenty rats with a peroneal to tibial ETS neurorrhaphy were divided into two groups: (1) control group and (2) IL-10 group with intrafascicular application of 0.125 µg/100 µl IL-10. Survival time was 8 weeks. Nerve conduction velocities (NCVs) and motor function were analyzed and histomorphological evaluation with measurement of intraneural collagen level, axon count, total nerve area, and myelination index followed. Evaluation of motor function and nerve conduction did not show any statistical differences. Histological analyses revealed thicker myelin sheaths and higher myelination index in the IL-10 group (p < 0.001). Axon count showed no difference. The IL-10 group revealed lower collagen levels (p < 0.001). Comparison of total nerve area showed no statistical significance. At this dose, IL-10 evaluated at 8 weeks was not significantly different than placebo in functional, NCVs, and most morphological measures. However, there was a significant difference in thicker myelin sheaths and higher myelination index and lower collagen levels. This suggests that future experiments of IL-10 at different doses or longer periods of evaluation would be of interest.