Age and Gender Estimation Using the Osseous Microanatomy: Original Research.

Aim: This study was performed with the idea of assessing age and gender utilizing differences in osseous microanatomy in human jawbones. Materials and Methods: The study was conducted retrospectively among human jawbone samples. Various morphometric assessments such as trabecular width, marrow space, and their corelation were studied. In the samples, variations among osteon numbers, differences in the shapes of the osteocytes of jawbones, and amount of inflammation in the bony areas were recorded. Results: It was noted in this study that mean values of the diameter of the Haversian canal and vessel density had a noteworthy increase in female jawbone samples. The amount of osteocytes in both female and male bone samples was also statistically significant in terms of the correlation coefficient. Conclusion: We concluded that more sensitive identification of human remains, that is, age and gender analysis, can be performed by histomorphometric evaluation of bone remains.

The validation of computed tomography derived radiodensity measurements of bone healing using histopathology.

BACKGROUND: The early identification of delayed bone healing or a non-union is vital for prompt treatment and superior patient outcomes. Current techniques rely heavily on operator skill for interpretation and hence their reliability and repeatability may be inconsistent. This study assessed the application of computed tomography (CT) derived densiometric measurements as a quantitative tool for the assessment of bone healing. METHODS: This prospective, longitudinal, method comparison study was performed using a recognised sheep tibial ostectomy model. Secondary bone healing was assessed at 2, 4, 6, and 8 weeks after the ostectomy was performed. CT densiometric measures of bone healing (Hounsfield units) were taken of the cis, trans, cranial and caudal cortices relative to the bone plate, with histological measurements (percentage of ossification) sourced from the same areas. Cis cortical densiometric data points were excluded from analysis due to significant beam hardening artefact from the bone plate (P < 0.001). A univariable linear regression was performed on the remaining data using averaged radiodensity (independent variable) and histomorphometric (dependent variable) measurements. RESULTS: The two measurements were significantly correlated (R2 = 0.623, P = 0.020) with a clear positive trend identified. CONCLUSION: This study suggests that radiodensity measurements may be a useful diagnostic and management tool for the monitoring of indirect bone healing.

Identifying primary tumor site of origin for liver metastases via a combination of handcrafted and deep learning features.

Liver is one of the most common sites for metastases, which can occur on account of primary tumors from multiple sites of origin. Identifying the primary site of origin (PSO) of a metastasis can help in guiding therapeutic options for liver metastases. In this pilot study, we hypothesized that computer extracted handcrafted (HC) histomorphometric features can be utilized to identify the PSO of liver metastases. Cellular features, including tumor nuclei morphological and graph features as well as cytoplasm texture features, were extracted by computer algorithms from 175 slides (114 patients). The study comprised three experiments: (1) comparing and (2) fusing a machine learning (ML) model trained with HC pathomic features and deep learning (DL)-based classifiers to predict site of origin; (3) identifying the section of the primary tumor from which metastases were derived. For experiment 1, we divided the cohort into training sets composed of primary and matched liver metastases [60 patients, 121 whole slide images (WSIs)], and a hold-out validation set (54 patients, 54 WSIs) composed solely of liver metastases of known site of origin. Using the extracted HC features of the training set, a combination of supervised machine classifiers and unsupervised clustering was applied to identify the PSO. A random forest classifier achieved areas under the curve (AUCs) of 0.83, 0.64, 0.82, and 0.64 in classifying the metastatic tumor from colon, esophagus, breast, and pancreas on the validation set. The top features related to nuclear and peri-nuclear shape and textural attributes. We also trained a DL network to serve as a direct comparison to our method. The DL model achieved AUCs for colon: 0.94, esophagus: 0.66, breast: 0.79, and pancreas: 0.67 in identifying PSO. A decision fusion-based strategy was deployed to fuse the trained ML and DL classifiers and achieved slightly better results than ML or DL classifier alone (colon: 0.93, esophagus: 0.68, breast: 0.81, and pancreas: 0.69). For the third experiment, WSI-level attention maps were also generated using a trained DL network to generate a composite feature similarity heat map between paired primaries and their associated metastases. Our experiments revealed that epithelium-rich and moderately differentiated tumor regions of primary tumors were quantitatively similar to paired metastatic tumors. Our findings suggest that a combination of HC and DL features could potentially help identify the PSO for liver metastases while at the same time also potentially identify the spatial sites of origin for the metastases within primary tumors.

Comparative Histomorphometric Analysis of Bone Regeneration According to Bone Graft Type.

PURPOSE: To use histomorphometric analysis to evaluate bone reconstruction in rabbit calvaria with autogenous bone, anorganic bovine bone, undecalcified human tooth bone (UdTB), and decalcified human tooth bone (dTB) grafts. MATERIALS AND METHODS: Extracted human teeth were crushed, and tooth bone with and without decalcification was prepared. Bony defects were made in 10 rabbit calvaria and allocated to one of the following four groups: group 1, in which UdTB was grafted; group 2, in which dTB was grafted; group 3, in which anorganic bovine bone was grafted; group 4, in which autogenous bone was grafted. The rabbits were sacrificed at 2 or 8 weeks postoperatively, and histomorphometric comparison was performed. RESULTS: Histologically, new bone formation was observed at the defect margin and around all graft materials. The dTB group revealed significantly greater new bone areas at 2 and 8 weeks compared to the UdTB group and the anorganic bovine bone group (P < .05). The dTB group revealed no significant difference in the new bone area at 2 weeks but revealed significantly less new bone area at 8 weeks compared to the autogenous bone group (P < .05). The dTB group also revealed significantly less graft material area compared to the anorganic bovine bone group at 8 weeks (P < .05). The autogenous bone group revealed significantly less graft material area and significantly greater bone marrow area compared to other groups at 8 weeks (P < .05). CONCLUSIONS: Grafting with dTB resulted in better bone regeneration than UdTB and anorganic bovine bone grafting at 8 weeks and addresses the potential disadvantages of autogenous bone grafting.

Morphological Reconstruction of a Critical-Sized Bone Defect in the Maxillofacial Region Using Modified Chitosan in Rats with Sub-Compensated Type I Diabetes Mellitus.

It is known that complexes based on natural polysaccharides are able to eliminate bone defects. Prolonged hyperglycemia leads to low bone regeneration and a chronic inflammatory response. The purpose of this study was to increase the efficiency of early bone formation in a cavity of critical size in diabetes mellitus in the experiment. The polyelectrolyte complex contains high-molecular ascorbate of chitosan, chondroitin sulfate, sodium hyaluronate, heparin, adgelon serum growth factor, sodium alginate and amorphous nanohydroxyapatite (CH-SA-HA). Studies were conducted on five groups of white female Wistar rats: group 1-regeneration of a bone defect in healthy animals under a blood clot; group 2-regeneration of a bone defect under a blood clot in animals with diabetes mellitus; group 3-bone regeneration in animals with diabetes mellitus after filling the bone cavity with a collagen sponge; group 4-filling of a bone defect with a CH-SA-HA construct in healthy animals; group 5-filling of a bone defect with a CH-SA-HA construct in animals with diabetes mellitus. Implantation of the CH-SA-HA construct into bone cavities in type I diabetic rats can accelerate the rate of bone tissue repair. The inclusion of modifying polysaccharides and apatite agents in the construction may be a prospect for further improvement of the properties of implants.

Whole slide imaging (WSI) scanner differences influence optical and computed properties of digitized prostate cancer histology.

Purpose: Digital pathology is becoming an increasingly popular area of advancement in both research and clinically. Pathologists are now able to manage and interpret slides digitally, as well as collaborate with external pathologists with digital copies of slides. Differences in slide scanners include variation in resolution, image contrast, and optical properties, which may influence downstream image processing. This study tested the hypothesis that varying slide scanners would result in differences in computed pathomic features on prostate cancer whole mount slides. Design: This study collected 192 unique tissue slides from 30 patients following prostatectomy. Tissue samples were paraffin-embedded, stained for hematoxylin and eosin (H&E), and digitized using 3 different scanning microscopes at the highest available magnification rate, for a total of 3 digitized slides per tissue slide. These scanners included a (S1) Nikon microscope equipped with an automated sliding stage, an (S2) Olympus VS120 slide scanner, and a (S3) Huron TissueScope LE scanner. A color deconvolution algorithm was then used to optimize contrast by projecting the RGB image into color channels representing optical stain density. The resulting intensity standardized images were then computationally processed to segment tissue and calculate pathomic features including lumen, stroma, epithelium, and epithelial cell density, as well as second-order features including lumen area and roundness; epithelial area, roundness, and wall thickness; and cell fraction. For each tested feature, mean values of that feature per digitized slide were collected and compared across slide scanners using mixed effect models, fit to compare differences in the tested feature associated with all slide scanners for each slide, including a random effect of subject with a nested random effect of slide to account for repeated measures. Similar models were also computed for tissue densities to examine how differences in scanner impact downstream processing. Results: Each mean color channel intensity (i.e., Red, Green, Blue) differed between slide scanners (all P<.001). Of the color deconvolved images, only the hematoxylin channel was similar in all 3 scanners (all P>.05). Lumen and stroma densities between S3 and S1 slides, and epithelial cell density between S3 and S2 (P>.05) were comparable but all other comparisons were significantly different (P<.05). The second-order features were found to be comparable for all scanner comparisons, except for lumen area and epithelium area. Conclusion: This study demonstrates that both optical and computed properties of digitized histological samples are impacted by slide scanner differences. Future research is warranted to better understand which scanner properties influence the tissue segmentation process and to develop harmonization techniques for comparing data across multiple slide scanners.

Cone-Beam Computed Tomography and Histological Findings for Socket Preservation Techniques Using Different Grafting Materials: A Systematic Review.

OBJECTIVE: Socket preservation techniques have been used to maintain the ridge dimension following tooth extraction. The materials used influence the quality and quantity of newly formed bone. Therefore, the aim of this article was to systematically review the literature reporting both histological and radiographic outcomes of socket preservation techniques after tooth extraction in human subjects. MATERIAL AND METHOD: A systematic electronic search was performed in the electronic databases. English language clinical studies that were published between 2017 and 2022 and included both histological and radiographic findings for the test and control groups. Our primary search produced 848 articles, and of these, 215 were duplicate studies. A total of 72 articles were then eligible for full-text reading. RESULTS: The review included eight studies that met its inclusion criteria. Three outcomes were compared in the included studies. The percentage of newly formed bone ranged from 21.34 ± 9.14% to more than 50% of new bone formation. The materials that showed more than 50% of newly formed bone formation were demineralized dentin graft, platelet-rich fibrin, freeze-dried bone allograft, corticocancellous porcine, and autogenous bone. Four Studies did not report the percentage of the residual graft materials, while those who reported showed a variable range of a minimum 1.5% to more than 25%. One study did not report the changes in horizontal width at the follow-up period, while other studies ranged from 0.6 mm to 10 mm. CONCLUSION: Socket preservation represents an efficient technique to preserve the ridge contour with satisfactory newly formed bone in the augmented site and maintaining the vertical and horizontal dimensions of the ridge.

Healing of particulate allografts mixed with platelet concentrates in ridge preservation and sinus lift: a prospective histomorphometric study.

The objective of the present study was to evaluate the bone quality of sinus and alveolar grafts following filling with particulate allogenous bone (DFDBA 300-500µm) and platelet concentrate (platelet-rich fibrin, PRF). A prospective interventional clinical study was carried out. A total of 40 bone cores, 2mm in diameter, were taken from 21 patients: 22 from grafted alveoli, 7 from grafted sinus sites, and 11 from native bone used as a control. Fixed, paraffin-embedded samples were subjected to histological staining with hematoxylin-eosin and Masson's trichrome. Bone maturity of the samples was evaluated by two independent operators using histomorphometric analysis. There existed a greater proportion of lamellar neoformed bone than woven neoformed bone as the healing time increased. Moreover, there was also an increasing proportion of newly formed bone in the grafted sockets as a function of healing time (average: 41.22% ≤ 5 months, 55.89% Ëƒ 5 months). Resorption of DFDBA particles also appears to be correlated with healing time in the grafted socket (average: 15.43 ≤ 5 months, 13.72% Ëƒ 5 months). In conclusion, performing sinus lift and alveolar socket preservation techniques using DFDBA and PRF results in high quality, mature bone tissue according to histological criteria.

Effect of commercially pure titanium implant coated with calcium carbonate and nanohydroxyapatite mixture on osseointegration.

In this research, rabbit femurs were implanted with CP Ti screws coated with a combination of CaCO3 and nanohydroxyapatite, and the effect on osseointegration was assessed using histological and histomorphometric examination at 2 and 6 weeks. CaCO3 and nanohydroxyapatite were combined with the EPD to coat the surfaces of the CP Ti screws. The femurs of five male rabbits were implanted with coated and uncoated implant screws. Healing time was divided into two groups (2 and 6 weeks). After 2 and 6 weeks of implantation, the histological examination revealed an increase in the growth of bone cells for coated screws, and the histomorphometric analysis revealed an increase in the percentage of new bone formation (after 6 weeks, 5.08% for coated implants and 3.66% for uncoated implants). In addition, the uncoated implant, the CP Ti implant coated with a combination of CaCO3 and nanohydroxyapatite, stimulated early bone development after two weeks and mineralization and maturation after six weeks.

Effect of magnetized water on blood indices and histomorphometric parameters of intestinal sections of Japanese quail.

A two-way experimental design was used to demonstrate the physiological effects of magnetized water and sex on blood indices and histomorphometric parameters of Japanese quail intestine sections. Red blood cell count (RBCs), packed cell volume (PCV), hemoglobin concentration (Hb), thrombocytes, white blood cell count (WBCs), and WBC differentiation were investigated. A total of 450 unsexed Japanese quail were randomized into three groups (45/replicate; 3 replicates; 135/group). As a monitoring group, the first group was given untreated tap water to drink. The two others were consumed magnetized water that were subjected to an electrical magnetic field with a power of 1 Tesla (10,000 Gauss) and 2 Tesla (20,000 Gauss), respectively. The treatments had a significant (p ≤ 0.05) effect on thrombocytes and Hb. Sex showed significant (p ≤ 0.05) differences for RBCs and PCV at 42 days of age. At different ages, significant effects were observed on histomorphometric parameters of the Japanese quail intestinal tract. It may be inferred that the influence of magnetized water, up to 1 Tesla, was positive on the haematological and histomorphometric parameters of the Japanese quail intestinal tract by augmenting the haematological measurements, which were within a normal range and increasing the surface area of the villus.

A Bone Histomorphometric Analysis of Hypophosphatasia-related Osteoporosis after Teriparatide Treatment.

A 79-year-old man was admitted with a compression fracture of the first lumbar vertebra. His alkaline phosphatase (ALP) level was 35 IU/L, and his dual energy X-ray absorptiometry T score was -3.7 standard deviations, indicating osteoporosis. A genetic analysis showed a mutation of the alkaline phosphatase biomineralization-associated gene encoding tissue-nonspecific alkaline phosphatase. Hypophosphatasia-related osteoporosis was diagnosed. Alendronate, teriparatide, and minodronate were administered in that order. The ALP level increased during teriparatide use. A bone biopsy performed after three years of teriparatide treatment showed that cancellous bone was adynamic. In cortical bone, tetracycline double-labeling indicates enhanced bone formation. Teriparatide may thus be a viable treatment option even in patients with hypophosphatasia.

Assessment of 28-day oral exposure to Pueraria candollei var. mirifica (Fabaceae) roots on pituitary-ovarian axis function and selected metabolic parameters in ovary-intact rats.

Pueraria candollei var. mirifica (Fabaceae) root (PMR) has recently been developed as a potential selective estrogen receptor modulator (SERM) in menopausal women. Nowadays, many premenopausal women also take dietary PMR supplements, however, the exact biological effects of PMR have not been evaluated. This study included the application of the OECD guideline 407 for the assessment of 28-day oral exposure to PMR on pituitary-ovarian (PO) axis function and metabolic parameters in the premenopausal rat model. Ovary-intact adult rats were orally administrated with 10, 100, 750, 1000, and 1500 mg/kg body weight (BW)/day of PMR powder. The positive estrogenic group was given 2 mg 17ß-estradiol (E2)/kg BW/day. Serum levels of reproductive hormones, lipid and thyroid parameters, estrous cycle determination, and histomorphometric and histopathological evaluations of the anterior pituitary, ovary, uterus, vagina, mammary gland, and liver were investigated. PMR displayed neutral effects on uterine, vaginal, and body weights, and circulating E2 and prolactin levels. PMR exerted E2-like effects by i) reducing ovarian and increasing hepatic weights, ii) decreasing serum gonadotropins, iii) lowering serum lipids without altering thyroid parameters, iv) increasing the prevalence of abnormal estrous cycles with prolonged estrus, v) increasing nuclear diameter of anterior pituitary cells, vi) decreasing ovarian size and follicular numbers and increasing follicular degeneration, vii) thickening of uterine myometrium and luminal epithelium, and vaginal epithelium, and viii) induction of mammary alveolar hyperplasia and ductal secretion. Unlike E2, the appearance of very small numbers of focal microvesicular steatosis in hepatocytes demonstrated mild toxicity at high PMR doses. This is the first report that high-dose PMR exerted actions exactly like E2 on gonadotrope-ovarian axis function and histology, lipid, and thyroid parameters without affecting uterine and vaginal growth in ovary-intact rats according to OECD guidelines.

Which Histometric Analysis Approach Is More Reliable for Assessing Histological Bone Tissue Samples?

This study aims to evaluate the grid of Merz and ImageJ methods for histometric quantification, verifying which is more reliable and defining which is most suitable based on the time required to perform. Thirty histological samples of maxillary sinuses grafted with xenografts were evaluated using an optical light microscope attached to an image capture camera and connected to a microcomputer. The images were digitalized and recorded as a TIFF image, and the new bone formation was evaluated using the grid of Merz and ImageJ. The Bland-Altman analysis was used to identify the agreement between the methods and determine suitable future research options. The timing of the quantification was also performed to identify a possible advantage. The mean value for the quantification analysis timing for the grid of Merz was 194.9 ± 72.0 s and for ImageJ was 871.7 ± 264.4, with statistical significance between the groups (p = 0.0001). The Bland-Altman analysis demonstrated a concordance between the methods, due to the bias being next to the maximum concordance (-1.25) in addition to the graphic showing the scattering points next to the mean of differences and inside of limits of agreement. Thus, it was demonstrated that the grid of Merz presents reliable outcomes and advantages over the ImageJ methodology regarding the time spent to contour the areas of interest.

Probability-based approach for characterization of microarchitecture and its effect on elastic properties of trabecular bone.

Trabecular bone is a random cellular solid with an interconnected network of plate-like and rod-like components. However, the structural randomness and complexity have hindered rigorous mathematical modeling of trabecular bone microarchitecture. Recent advancements in imaging processing techniques have enabled us to define the size, orientation, and spatial location of individual trabecular plates and rods in trabecular bone. Based on the essential information, this study proposed a probability-based approach to define the size, orientation, and spatial distributions of trabecular plates and rods for trabecular bone cubes (N = 547) acquired from six human cadaver proximal femurs. Using two groups of probability-based parameters, it was attempted to capture microarchitectural details, which could not be captured by the existing histomorphometric parameters, but crucial to the elastic properties of trabecular bone. The elastic properties of the trabecular bone cubes in three principal axes were estimated using microCT based finite element (FE) simulations. Based on the results of multivariate multiple regression modeling, the efficacy of the two groups of probability-based parameters in prediction of the elastic properties was verified in comparison with that of the existing histomorphometric parameters (BV/TV, Tb.Th, Tb.Sp, DA, EF.Med, and Conn.D). The results indicated that the regression models trained using the probability-based parameters had a comparable and even better accuracy (rMSE = 0.621 and 0.548) than that of the histomorphometric parameters (rMSE = 0.647). More importantly, the probability-based parameters could provide more insights into some unexplored microarchitectural features, such as individual trabecular size, orientation, and spatial distributions, which are also critical to the elastic properties of trabecular bone.

Age-Related Changes in Calcitonin-Producing Thyroid C-Cells of Male Wistar Rats.

Thyroid C-cells secrete the hormone calcitonin (CT) which acts as an inhibitor of bone resorption. Our aim was to examine the age-related changes in the structure and function of CT-producing C-cells, using histomorphometric, ultrastructural, and biochemical analyses. We used young adult (3-months-old), middle-aged (16-months-old), and old (24-months-old) male rats. The peroxidase-antiperoxidase method was applied for localization of CT. Stereological analysis was performed using the newCAST stereological software package. Serum samples were analyzed for the determination of CT, testosterone (T), calcium (Ca2+), and phosphorus (P). We found a significant increase in the volume density (Vv) of C-cells in both older groups (p < 0.05). The percentage of smaller volume range C-cells increased (p < 0.0001), while the proportion of greater volume range C-cells decreased (p < 0.05) with ageing. Ultrastructural analysis revealed a larger number of secretory granules in older rats. Serum CT increased (p < 0.001), while serum T and P were reduced (p < 0.01) in older rats. Serum Ca2+ was lower (p < 0.0001) in middle-aged rats compared to young adults. We revealed a 20% incidence of C-cell hyperplasia in older rats and one case of medullary thyroid carcinoma in an old rat. Our findings indicate that the ageing process causes significant histomorphometric changes at the thyroid C-cell level.

Seasonal Variations in the Digestive Tract of the Little Owl, Athene noctua: Anatomical, Histological, and Scanning Electron Microscopical Studies.

The digestive tract of the little owl, Athene noctua (Strigiformes: Strigidae), is described in two different seasons. The digestive tract of this bird follows the basic model for that of a predatory bird. The cervical esophagus is not expanded to form a crop. The internal surface of the esophagus forms numerous longitudinal folds provided with numerous mucous glands. These longitudinal folds increase in number and vary in depth posteriorly. The folds of the proventriculus are composed of simple branched tubular glands. The ventriculus is lined by a thin layer of koilin. The number of goblet cells gradually increases from the duodenum to the rectum, and the lymphatic tissue diffuses within the lamina propria. The esophageal glands secrete acid mucopolysaccharides, while the gastric glands of the stomach, the goblet cells, and crypts of Lieberkühn secrete acid mucopolysaccharides. Proteins were observed in the different histological structures of the digestive tract. Morphometric and histometric studies showed differences between summer and winter in the esophagus and glandular stomach (especially in winter), but no seasonal differences were seen in the muscular stomach, or small and large intestines.

Osteogenic Induction with Silicon Hydroxyapatite Using Modified Autologous Adipose Tissue-Derived Stromal Vascular Fraction: In Vitro and Qualitative Histomorphometric Analysis.

Large bone defects requiring invasive surgical procedures have long been a problem for orthopedic surgeons. Despite the use of autologous bone grafting, satisfactory results are often not achieved due to associated limitations. Biomaterials are viable alternatives and have lately been used in association with Stromal Vascular Fraction (SVF), stem cells, and signaling factors for bone tissue engineering (BTE). The objective of the current study was to assess the biocompatibility of Silicon Hydroxyapatite (Si-HA) and to improve osteogenic potential by using autologous adipose-derived SVF with Si-HA in a rabbit bone defect model. Si-HA granules synthesized using a wet precipitation method were used. They were characterized using scanning electron microscopy (SEM), Fourier transform infrared (FTIR), and X-ray diffraction (XRD). A hemolysis assay was used to assess the hemolytic effects of Si-HA, while cell viability was assessed through Alamar Blue assay using MC3T3 mouse osteoblasts. The osteogenic potential of Si-HA both alone and with enzymatically/non-enzymatically-derived SVF (modified) was performed by implantation in a rabbit tibia model followed by histomorphometric analysis and SEM of dissected bone after six weeks. The results showed that Si-HA granules were microporous and phase pure and that the addition of Silicon did not influence Si-HA phase composition. Si-HA granules were found to be non-hemolytic on the hemolysis assay and non-toxic to MC3T3 mouse osteoblasts on the Alamar Blue assay. Six weeks following implantation Si-HA showed high biocompatibility, with increased bone formation in all groups compared to control. Histologically more mature bone was formed in the Si-HA implanted along with non-enzymatically-derived modified SVF. Bone formation was observed on and around Si-HA, reflecting osseointegration. In conclusion, Si-HA is osteoconductive and promotes osteogenesis, and its use with SVF enhances osteogenesis.

Early unhealthy eating habits underlie morpho-functional changes in the liver and adipose tissue in male rats.

Early-life consumption of high-fat and sugar-rich foods is recognized as a major contributor for the onset of metabolic dysfunction and its related disorders, including diabetes and nonalcoholic fatty liver disease. The lifelong impact of early unhealthy eating habits that start at younger ages remains unclear. Therefore, to better understand the effects of diet, it is essential to evaluate the structural and functional changes induced in metabolic organs and potential mechanisms underlying those changes. To investigate the long-term effects of eating habits, young male rats were exposed to high-sugar and high-energy diets. After 14 weeks, body composition was assessed, and histopathological changes were analyzed in the liver and adipose tissue. Serum biochemical parameters were also determined. Expression of inflammatory markers in the liver was evaluated by immunohistochemistry. Our results revealed that serum levels of glucose, creatinine, aspartate transaminase (AST), alanine transaminase (ALT), and lipid profile were increased in rats red high-sugar and high-energy diets. Histopathological alterations were observed, including abnormal hepatocyte organization and lipid droplet accumulation in the liver, and abnormal structure of adipocytes. In both unhealthy diet groups, hepatic expression of Toll-like receptor 4 (TLR4), cyclooxygenase 2 (COX-2), and E-selectin were increased, as well as a biomarker of oxidative stress. Together, our data demonstrated that unhealthy diets induced functional and structural changes in the metabolic organs, suggesting that proinflammatory and oxidative stress mechanisms trigger the hepatic alterations and metabolic dysfunction.

The Effect of Secretome, Xenogenic Bone Marrow-Derived Mesenchymal Stem Cells, Bone Morphogenetic Protein-2, Hydroxyapatite Granule and Mechanical Fixation in Critical-Size Defects of Rat Models.

Background: Recent studies have shown that human bone marrow-derived mesenchymal stem cells (hBM-MSCs) have several drawbacks in treating critical-sized bone defect (CSD). Secretome may offer considerable advantages over living cells in terms of potency, manufacturing and storing easiness, and potential as a ready-to-go osteoinductive agent. However, thus far, there are no studies regarding the efficacy of secretome in bone healing. The objective of this study is to investigate the effect of the secretome in rat models with CSD. Methods: This was an experimental study with post-test only control group design using 60 skeletally mature Sprague Dawley rat which was divided evenly into 5 treatment groups (MSC only, Secretome only, MSC + Secretome, MSC + Secretome + BMP-2, Control group using Normal Saline). We used Bone Marrow derived MSC in this research. The critical-sized bone defect was created by performing osteotomy and defect was treated according to the groups. Rats were sacrificed on 2nd and 4th week and we measured the radiological outcome using Radiographic Union Score for Tibia (RUST) and histomorphometric (callus, osseous, cartilage, fibrous, and void area) evaluation using Image J. Results: There was no difference in the weight of rats between groups before and after the intervention. RUST score in all intervention group is significantly higher than the control group, however, the MSC-only group was not statistically significant higher than the control group. There is no statistically significant difference in RUST Score between intervention groups.Histomorphometric evaluation showed that total callus formation is the widest in the MSC+Secretome+BMP-2 combination group while the osseous area is found highest on the secretome-only group. Conclusion: Secretome, whether used solely or combined with BM-MSC and BMP-2, is a novel, potent bone-healing agent for CSD in rat models.

Effects of Creatine Supplementation on Histopathological and Biochemical Parameters in the Kidney and Pancreas of Streptozotocin-Induced Diabetic Rats.

Diabetes mellitus (DM) is a worldwide health concern, and projections state that cases will reach 578 million by 2030. Adjuvant therapies that can help the standard treatment and mitigate DM effects are necessary, especially those using nutritional supplements to improve glycemic control. Previous studies suggest creatine supplementation as a possible adjuvant therapy for DM, but they lack the evaluation of potential morphological parameters alterations and tissue injury caused by this compound. The present study aimed to elucidate clinical, histomorphometric, and histopathological consequences and the cellular oxidative alterations of creatine supplementation in streptozotocin (STZ)-induced type 1 DM rats. We could estimate whether the findings are due to DM or the supplementation from a factorial experimental design. Although creatine supplementation attenuated some biochemical parameters, the morphological analyses of pancreatic and renal tissues made clear that the supplementation did not improve the STZ-induced DM1 injuries. Moreover, creatine-supplemented non-diabetic animals were diagnosed with pancreatitis and showed renal tubular necrosis. Therefore, even in the absence of clinical symptoms and unaltered biochemical parameters, creatine supplementation as adjuvant therapy for DM should be carefully evaluated.