The Effects of NPY1 Receptor Antagonism on Intervertebral Disc and Bone Changes in Ovariectomized Rats.

STUDY DESIGN: Basic science. OBJECTIVE: To compare the effects of a neuropeptide Y1 receptor antagonist (NPY-1RA) to estrogen on maintaining vertebral bone microarchitecture and disc height in a rat model of menopause. METHODS: This study was an institutional animal care approved randomized control study with 104 ovariectomized rats and 32 intact control animals. Comparison of disc height, trabecular bone, body weights, circulating levels of NPY and estrogen, and distribution of Y1 receptors in the intervertebral disc in an established rodent osteoporotic model were made at baseline and after 2, 4, and 8 weeks after receiving either an implant containing estrogen or an antagonist to the neuropeptide Y1 receptor. Data was compared statistically using One-way analysis of variance. RESULTS: Circulating levels of estrogen increased and NPY decreased following estrogen replacement, with values comparable to ovary-intact animals. NPY-1RA-treated animals had low estrogen and high NPY circulating levels and were similar to ovariectomized control rats. Both NPY-1RA and estrogen administration were able reduce, menopause associated weight gain. NPY-1RA appeared to restore bone formation and maintain disc height, while estrogen replacement prevented further bone loss. CONCLUSION: NPY-1RA in osteoporotic rats activates osteoblast production of bone and decreased marrow and body fat more effectively than estrogen replacement when delivered in similar concentrations. Annulus cells had NPY receptors, which may play a role in disc nutrition, extracellular matrix production, and pain signaling cascades.

The role of the msaABCR operon in implant-associated chronic osteomyelitis in Staphylococcus aureus USA300 LAC.

BACKGROUND: The msaABCR operon regulates several staphylococcal phenotypes such as biofilm formation, capsule production, protease production, pigmentation, antibiotic resistance, and persister cells formation. The msaABCR operon is required for maintaining the cell wall integrity via affecting peptidoglycan cross-linking. The msaABCR operon also plays a role in oxidative stress defense mechanism, which is required to facilitate persistent and recurrent staphylococcal infections. Staphylococcus aureus is the most frequent cause of chronic implant-associated osteomyelitis (OM). The CA-MRSA USA300 strains are predominant in the United States and cause severe infections, including bone and joint infections. RESULTS: The USA300 LAC strain caused significant bone damage, as evidenced by the presence of severe bone necrosis with multiple foci of sequestra and large numbers of multinucleated osteoclasts. Intraosseous survival and biofilm formation on the K-wires by USA300 LAC strains was pronounced. However, the msaABCR deletion mutant was attenuated. We observed minimal bone necrosis, with no evidence of intramedullary abscess and/or fibrosis, along reduced intraosseous bacterial population and significantly less biofilm formation on the K-wires by the msaABCR mutant. microCT analysis of infected bone showed significant bone loss and damage in the USA300 LAC and complemented strain, whereas the msaABCR mutant's effect was reduced. In addition, we observed increased osteoblasts response and new bone formation around the K-wires in the bone infected by the msaABCR mutant. Whole-cell proteomics analysis of msaABCR mutant cells showed significant downregulation of proteins, cell adhesion factors, and virulence factors that interact with osteoblasts and are associated with chronic OM caused by S. aureus. CONCLUSION: This study showed that deletion of msaABCR operon in USA300 LAC strain lead to defective biofilm in K-wire implants, decreased intraosseous survival, and reduced cortical bone destruction. Thus, msaABCR plays a role in implant-associated chronic osteomyelitis by regulating extracellular proteases, cell adhesions factors and virulence factors. However additional studies are required to further define the contribution of msaABCR-regulated molecules in osteomyelitis pathogenesis.

Collagen-Elastin-Like Polypeptide-Bioglass Scaffolds for Guided Bone Regeneration.

The goals of this study are to evaluate the ability of the multicomponent collagen-elastin-like polypeptide (ELP)-Bioglass scaffolds to support osteogenesis of rat mesenchymal stem cells (rMSCs), demonstrate in vivo biocompatibility by subcutaneous implantation in Sprague-Dawley rats, monitor degradation noninvasively, and finally assess the scaffold's ability in healing critical-sized cranial bone defects. The collagen-ELP-Bioglass scaffold supports the in vitro osteogenic differentiation of rMSCs over a 3 week culture period. The cellular (rMSC-containing) or acellular scaffolds implanted in the subcutaneous pockets of rats do not cause any local or systemic toxic effects or tumors. The real-time monitoring of the fluorescently labeled scaffolds by IVIS reveals that the scaffolds remain at the site of implantation for up to three weeks, during which they degrade gradually. Micro-CT analysis shows that the bilateral cranial critical-sized defects created in rats lead to greater bone regeneration when filled with cellular scaffolds. Bone mineral density and bone microarchitectural parameters are comparable among different scaffold groups, but the histological analysis reveals increased formation of high-quality mature bone in the cellular group, while the acellular group has immature bone and organized connective tissue. These results suggest that the rMSC-seeded collagen-ELP-Bioglass composite scaffolds can aid in better bone healing process.

SODIUM BICARBONATE REMEDIATION OF ANTHROPOGENIC CONTAMINATION OF WATER AT THE GBNERR IN MISSISSIPPI.

Grand Bay National Estuarine Research Reserve (GBNERR) is an important ecosystem in the Mississippi Gulf Coast. The GBNERR may be a potential source for contamination with anthropogenic bacterial pathogens that may play a significant role in the causation of waterborne human diseases. The objective of this study was to evaluate the interaction of physicochemical and microbiological water quality parameters at the GBNERR, determine quantitative levels and establish the potential for remediation of post-contamination of water and seafood by human fecal pollution from anthropogenic sources at the reserve. Water samples were collected aseptically from Bayous Heron, Cumbest, Point Aux Chenes Bay and Bangs Lake (Pine-O-Pine). Physicochemical parameters were determined using standard protocols. Eight bacterial species including Campylobacter were concentrated from water samples by membrane filtration. Water samples were tested for the presence of traditional indicator microorganisms including: heterotrophic (HPC), total coliforms (TC), fecal coliforms (FC) and enterococcus (ENT) in CFU/ml concentrations. Mean values of temperature, specific conductivity, dissolved oxygen and pH were within acceptable levels in comparison to MDEQ, USEPA and the USGS standards during the time of investigation. However, the values of turbidity in Grand Bay water exceeded USEPA recommended levels in several occasions during the investigation. Data from this study indicates significant variability (p < 0.0001) in mean bacteria concentrations between sites. The data also indicates significant impact of Sodium bicarbonate treatment in the remediation of post contamination and survival of pathogens from the GBNERR Bayous Heron, Cumbest and Pine-O-Pine when compared with control findings. The interaction of physicochemical and microbiological parameters of water through external chemical manipulation by Sodium bicarbonate may provide utility in the remediation of post-contamination with anthropogenic pathogens such as E. coli, Enterococci, Campylobacter, Vibrio, Giardia and Cryptosporidium. Presence of high numbers of indicator bacteria suggest public health concerns for oyster and shellfish consumers as well as other water contact activities. Hence, control strategies should be developed and implemented to prevent or remediate any future contamination of the GBNERR waters citing the economic impact of such contamination on shell fish fishing activities at the reserve.

THE IMPACT OF HYDROXYAPATITE SINTERING TEMPERATURES ON THE STRUCTURAL AND FUNCTIONAL CAPACITY OF EMBRYONAL LUNG FIBROBLASTS.

The specific aim of this investigation was to investigate the effects of hydroxyapatite (HA) sintered at different temperatures on the proliferation, morphology, and cellular alterations of Embryonal Lung Fibroblasts (MRC-5) cells in culture. Microcrystals of HA were prepared by following standard laboratory protocols. The calcined HA was then placed on an automatic Tyler sieve stack to select for particles between 1-40 µm in size. The calcined material was sintered at 700, 1000, 1200 and 1300 °c for 24 hours. The sintered material (1.0 grams of HA) was added to a total of 20 mg of L-lysine as a binder. Finally, this combination was cold-pressed into cylindrical form using a 3/8 inch die set at a compression load of 5000 kg to form a disk of HA. Calculated density of the devices was directly proportional to temperature of sintering. MRC-5 fibroblasts were obtained from the American Type Culture Collection (ATCC). Proliferation Assessment, morphological evaluations were conducted following standard lab protocols. Cells were analyzed for Maliondialdehyde (MDA) levels by utilizing thiobarbituric acid reactive substance (TBARS) measurements. Results of this study revealed that there was an initial (at 24, and 48 hours) increase in proliferation rate observed in wells containing HA sintered at 700, 1000, and 1300°C. There were no significant differences in proliferating cells among control and capsules sintered at 1200°C (P<0.05). However, at 72 hours there was 2-3 fold increase in cell number for the cells encountering 1200°C compared with control and experimental wells. Cellular membrane damage was evident in cells exposed to HA sintered at 700, 1000 and 1300°C, but not for control or HA 1200°C treated cells. Morphologically, the cells showed evidence of progressive fragmentations, cellular debris and lysis at 72 hours for cells in the HA sintered groups of 700, 1000 and 1300°C. HA sintered at 1200°C did not cause changes in cell morphology for the duration of the experiment. In conclusion, the results of this investigation suggest that sintering temperature is essential factor in the development of a high mechanical strength HA delivery systems. The results from this observation and our previous findings using different calcium phosphate devices suggest that the use of optimal HA density is crucial for material constructs to replace or repair tissue defects.

DIFFERENTIAL HISTOPATHOLOGICAL ASSESSMENT OF TESTICULAR FUNCTION UPON LONG-TERM EXPOSURE TO SUSTAINED DELIVERY OF TESTOSTERONE AND DIHYDROTESTOSTERONE.

The specific aim of this study was to assess, histopathologically, the seminiferous tubules area and germ layers upon the exposure to sustained delivery of testosterone (TE) and dihydrotestosterone (DHT) through tricalcium phosphate lysine devices (TCPL). A total of 140 adult Male rats (280-320 gm) were randomly divided into four equal groups. Groups 1 and 2 animals were implanted with TCPL loaded with 40 mg TE and 40 mg DHT, respectively. Groups 3 and 4 animals served as a sham group (empty devices), and a control group. For the treatment and sham groups, serum testosterone, LH and FSH levels were monitored at treatment periodic intervals of 1, 3, 6, 9, and 12 months. Histopathological evaluation of testicular issues (H&E) was conducted for each phase following standardized lab procedures. Results of this study indicated that: (i) endogenous testosterone and gonadotropin (LH/FSH) levels were suppressed to undetectable levels (<0.2 ng/mL) for a 1-year period by the sustained delivery of either TE or DHT compared to control and sham groups, (ii) a decrease in the luminal areas of seminiferous tubules retrieved from DHT treated group (P<0.05) in comparison with TE, (iii) an arrest of germ layers at the secondary spermatocyte at the end of the 3 month treatment with DHT and 1 month exposure to TE, and (iv) spermatogonia were intact and exhibited normal N/C ration for TE or DHT treated animals compared to sham and control groups. The overall conclusion obtained from this study indicated that TE loaded TCPL delivery devices can be used to induce azoospermia at an early phase and also provided evidence of the increased TE effectiveness to regulate fertility in an animal model.

ASSESSMENT OF ANIMAL MODELS AS SURROGATES FOR HUMAN TUMORS FROM THREE DIFFERENT ORGANS.

The compositional balance and distribution of trace metals/elements in various body tissues are essential key players in tissue and cellular homeostasis. Low Zn levels as well as overexpression of metalothioneins were implicated in the development and progression of various cancers including the prostate. Nonetheless, wider elemental profiles that relate cancer and normal phenotypes with regards to metal homeostasis were not well elucidated in the literature. Moreover, laboratory animals are currently used as accepted models for studying cancer but the level of their representation of actual cancer tissues was not clear. This study is attempting to assess the relevance of animal models currently in use, as surrogates for cancer and establish their relationship to actual normal and cancer tissues from humans. The major focus of this study was to investigate the differential relationship of metal concentrations and profiles in cancer and normal tissues from cadavers of humans and their comparison to established animal models representing organ cancers. The working hypothesis was that elemental/metal concentrations and profiles seen in post mortem will show significant differences between normal and cancer-derived tissues as well as between various tissue types in humans, rats, and dogs. This study also establishes critical elemental/metal profiles that may be relevant in providing correlations with the development of three major cancers. Normal human and tumor tissues of cadaverous lung, breast, and liver used in this study were obtained from US Biomax Company and relevant animal models (Sprague-dawley and Brown Norwegian rats as well as dogs; were obtained from Jackson Laboratories and the Mississippi State Veterinary Laboratory in Pearl, MS), to analyze for elements and test the hypothesis. Tissue samples were prepared using standardized digestion procedures necessary for use with the Inductively Coupled Plasma-Atomic Emission mass Spectrometry (ICP-MS) to determine the concentrations and profiles of 21 elements including Ag, Al, As, Ba, Ca, Cd, Co, Cr, Cu, Fe, Mg, Mn, Na, Ni, Pb, Sb, Se, Sr, Tl, V, and Zn. Our data supports the notion that metal/elemental homeostasis is essential for normal tissue function and that elemental variations in content, distributions, and ranking are tissue specific as well as carcinoma and species-specific. Analysis of data showed significant variations in elemental content and distribution profiles/ranking between animal models and actual human tissues consistent with the hypothesis. It is concluded that elemental homeostasis is essential for normal tissue function and that shifts in their distribution and content are essential in determining the use of animal models as surrogates for studying cancer. These results are promising and warrant further studies to confirm the relevance of animal models in relation to their use as pre-clinical tools for examining targeted cancer therapeutics.

Altered neuro-inflammatory gene expression in hippocampus in major depressive disorder.

Major Depressive Disorder (MDD) is a common psychiatric disorder for which available medications are often not effective. The high prevalence of MDD and modest response to existing therapies compels efforts to better understand and treat the disorder. Decreased hippocampal volume with increasing duration of depression suggests altered gene expression or even a decrease in neurogenesis. Tissue punches from the dentate gyrus were collected postmortem from 23 subjects with MDD and 23 psychiatrically-normal control subjects. Total RNA was isolated and whole transcriptome paired-end RNA-sequencing was performed using an Illumina NextSeq 500. For each sample, raw RNA-seq reads were aligned to the Ensembl GRCh38 human reference genome. Analysis revealed 30 genes differentially expressed in MDD compared to controls (FDR<0.05). Down-regulated genes included several with inflammatory function (ISG15, IFI44L, IFI6, NR4A1/Nur-77) and GABBR1 while up-regulated genes included several with cytokine function (CCL2/MCP-1), inhibitors of angiogenesis (ADM, ADAMTS9), and the KANSL1 gene, a histone acetyltransferase. Similar analyses of specific subsets of MDD subjects (suicide vs. non-suicide, single vs. multiple episodes) yielded similar, though not identical, results. Enrichment analysis identified an over-representation of inflammatory and neurogenesis-related (ERK/MAPK) signaling pathways significantly altered in the hippocampal dentate gyrus in MDD. Together, these data implicate neuro-inflammation as playing a crucial role in MDD. These findings support continued efforts to identify adjunctive approaches towards the treatment of MDD with drugs including anti-inflammatory and neuroprotective properties.

HISTOPATHOLOGICAL ANALYSIS OF THE F344 RAT LUNG UPON EXPOSURE TO RETENOIC ACID, OVALBUMIN, MOLD SPORES AND CITRAL.

The paradoxical role of retinoic acid (All Trans Retinoic Acid; ATRA) in the development of allergic and/or inflammatory complications in contrast to a therapeutic modality for lung pathology is not well understood or established in the literature. As well, the role of Citral (inhibitor of retinoid function; a non-toxic chemical that exists in two forms (diethyl; C1 or cis-trans dimethyl; C2), in the reversal of retinoic acid, ovalbumin and allergic mold spore pathophysiology is also not well ascertained under an in vivo setting. Therefore, it is hypothesized that exposure of F344 lung tissues to supra-physiologic levels of retinoic acid, ovalbumin and mold spores individually or in combination with each other will lead to inflammatory tissue pathology and that Citral 1 and 2 will reverse or ameliorate the related pathological damage to lung tissues. Even though ovalbumin and retinoic acid have been previously applied through intra-tracheal route in cancer prevention and immunological research, the objective of this study was to evaluate the histopathological implications of such exposure in vivo. This IACUC approved in vivo study used Fischer 344 rats (n = 80 ; 229 to 273g), which were randomly assigned to controls as well as ovalbumin and mold-sensitized treatment groups (0.80 mg/kg and 1×109 mold spores combined from 4 strains/100 µl intra-tracheal; all others were dosed by intra-peritoneal injection at days 1 and 7 with 80 mg/kg each of ATRA as well as 20 and 50 mg/kg each of Citrals 1 or 2 individually or in combination to represent all four chemicals and mold spores treatments. Positive and negative controls for each treatment were also included in the study. Animals were housed in rat cages at the JSU Research Animal Core Facilities and were placed on a 12:12 light-dark cycle. A standard rodent diet and water access were provided ad libidum. All animals were sacrificed on day 21 and lung tissues were processed for histopathology. Slides were prepared and were digitized for comparison of tissues pathology. Results showed that exposure of the F344 rats to ovalbumin and ATRA showed various levels of lung tissue damage that was ameliorated by Citral 2 in combination. Mold and ATRA exposure caused various levels of lung tissue damage that was reversed by C1 in combination with each other. Taken together, the study showed that there are variable pathologic inflammatory responses from the interaction of ovalbumin, Citrals, mold spores and retinoic acid, and that the addition of Citrals have reversed lung tissue pathologies. These findings warrants further investigation as to the actual role of these interactions in relation to acute/chronic lung disease and the possibility of reversing retinoid-mediated pathologies in the Fisher rat model.

IMPACT OF ATRA ON OVALBUMIN AND MOLD-SENSITIZED F344 RATS AND REVERSAL OF HEALTH-RELATED IMPLICATIONS BY CITRAL.

The role of retinoic acid (All Trans Retinoic Acid; ATRA) in the development of hypervitaminosis A pathophysiology is not well understood or established in the literature. As well, the role of Citral (inhibitor of retinoid function; a non-toxic chemical that exists in two forms (diethyl; C1 or cis-trans dimethyl; C2).) in the reversal of pathophysiological implications is also not ascertained under an in vivo setting. Therefore, it is hypothesized that ovalbumin exposure will sensitize the body to supra-physiologic levels of retinoic acid leading to a negative pathophysiological impact and that Citrals 1 and 2 will reverse or ameliorate the related damage to the body's pathophysiology. Even though ovalbumin and retinoic have been previously applied through intra-tracheal route in cancer prevention and immunological research, the objective of this study was to evaluate their interaction as a remedy for hypervitaminosis A. This IACUC approved in vivo study used Fischer 344 rats (n = 80 ;229 to 273g), which were randomly assigned to controls as well as ovalbumin and mold-sensitized treatment groups (0.80 mg/kg and 1X109 mold spores combined from 4 strains/100 µl intra-tracheal; all others were dosed by intra-peritoneal injection at days 1 and 7 with 80 mg/kg each of ATRA as well as 20 and 50 mg/kg each of Citrals 1 or 2 individually or in combination to represent all four chemicals and mold spores treatments.. Positive and negative controls for each treatment were also included in the study. Animals were housed in rat cages at the JSU Research Animal Core Facilities and were placed on a 12:12 light dark cycle. A standard rodent diet and water access were provided ad-libidum. Rat weights were recorded on day 1 and 21, all animals were sacrificed on day 21 and blood was collected and processed for hematological parameters. Results showed that even though C1 and C2 were not toxic individually, their combination at high dosing was lethal. Exposure of ovalbumin-sensitized rats to ATRA showed various levels of weight losses and negative hematological implications that were ameliorated by exposure to Citrals at various combinations with retinoic acid. Taken together, the study showed that there are variable pathophysiological responses from the interaction of ovalbumin, mold spores and retinoic acid and that Citrals were found to be individually effective in reversing health-related pathophysiologies. These findings warrants further investigations as to the actual role of these interactions in relation to acute pathophysiologic health implications and the possibility of reversing hypervitaminosis A-mediated health-related impacts.

Impact of Triple Combinations of Retinoic Acid, Mold Spores and Citral on the F344 Rat Lung Tissue Pathology.

The impact of retinoic acid (All Trans Retinoic Acid; ATRA) and Mold spores (MLD) in the development of lung pathology and in vivo tissue remodeling have not been well established in the literature. In addition, the role of citral (inhibitor of retinoid function) in the improvement of lung pathology has not been ascertained in animal studies. Therefore, it is hypothesized that ATRA and Mold (MLD) exposure will sensitize lung tissues leading to lung tissue pathology and that Citrals (C1 and C2) will reverse, ameliorate or improve the associated pathological damage to lung tissues. The study used an IACUC approved between-subject in vivo randomized split plot factorial design (F344 rat model; N=40). Animals were exposed to seven different treatments including untreated control, MLD, ATRA, Citrals (C1 and C2) and their MLD combinations (MLD+ ATRA+ C1, and MLD+ ATRA+ C2) by intra-peritoneal route. Rat weight and blood data were collected on Days 1 and 21, all animals were sacrificed on day 21, and lung tissues were processed for histopathology. Results from weight and blood data (ANOVA and Duncan) as well as from histopathological analyses supported the findings that exposure of F344 rats to MLD combinations with ATRA and Citrals showed various levels of lung tissue damage that were impacted by either C1 or C2 exposure. This promising study showed impressive responses on the interaction of MLD, Citrals, and ATRA as related to their impact on associated lung tissue pathologies.

Pathophysiological Effects of Thymoquinone and Epigallocatechin-3-Gallate on SK-OV-3 Ovarian Cancer Like Cell Line.

Ovarian cancer is the leading cause of mortality among the gynecologic cancers and is the 5th most common cancer among women. Recent studies have indicated that antioxidant exposure may slow the progression in major neoplastic diseases. The objective of this study was to investigate the pathophysiological effects of Thymoquinone (TQ) and Epigallocatechin-3-gallate (EGCG) using the SK-OV-3 cell line as a model. A total of 72 wells were plated with (10^5) SK-OV-3 ovarian cancer cells according to standard lab protocols. Each group was subdivided into 4 groups of 6 wells each. Group 1 served as control and groups 2, 3, and 4 were treated with TQ (16 µM), EGCG (3 µg/ml), and TQ + EGCG, respectively. Biomarker and morphological evaluations were performed following standard lab techniques. The results of the study revealed: (1) there were no differences in protein levels for all time periods (p<0.05); (2) an increase in nitric oxide following the administration of EGCG alone and in combination with TQ compared with TQ alone or untreated control cells with the difference being approximately three-fold higher and this difference was statistically different (p<0.05) at 24, 48, 72 hours; (3) glutathione levels were not statistically different at all time periods (p<0.05); and (4) the MDA levels showed no damage to the SK-OV-3 ovarian cancer cells. The results of this study provided the literature with more insights regarding manipulation of ovarian cancer behavior through potent antioxidants such as TQ and EGCG in culture.

Effects Of Polylactic Acid Coating and Compression Load on the Delivery of Protein and Steroid from HA Ceramic Devices.

Density variations, due to particle size and time and temperature of sintering, affect the delivery profile of substances from ceramic delivery devices. This investigation was conducted to study the effect of polylactic acid (PLA) impregnation on the porosity of hydroxyapatite (HA) capsules by studying the delivery rate of bovine serum albumin (BSA) and testosterone (TE). HA capsules were fabricated by cold compressing calcined particles at 1000, 3000, and 5000 lbs. Each group was subdivided into PLA-impregnated and non-PLA impregnated capsules. Each capsule was loaded with either 40 mg of TE (impregnated or nonimpregnated with PLA polymer) or BSA. Data obtained in this study suggest that: (1) PLA impregnation of HA ceramic capsules decreases the rate of release of drugs from the ceramic reservoir, (2) Physiochemical characteristics of the drugs to be delivered is an instrumental key in the sustained release profiles, (3) Compression load of HA reservoirs is a key factor in predicting the duration and level of sustained delivery, and (4) Polymer coating of HA ceramic capsules reduces the pore size, as well as, blocking some of the pores on the surface.

The Resultant Physiological Effects of Human Gingival Fibroblasts Exposed to Combinations of Dental Adhesives, Porphyromonas Gingivalis Lipopolysaccharide, and Cortisol or Nifedipine.

The needs of dental patients are becoming ever complex. This is largely due to the increasing number of systemic conditions that patients present with during routine or emergency dental visits, such as hypertension and angina pectoris. Medications that patients are prescribed have potential to cause effects on the oral tissues. There is a need to understand the interactions between the oral tissues and materials used to restore teeth. The objective of this study was to assess the physiological function of human gingival fibroblasts upon exposure to dental adhesives; polymethyl methacryate, OptiBond®, and Prime & Bond®; Porphyromonas gingivalis lipopolysaccharide and Cortisol or Nifedipine. An increase in metabolic activity was noted with the experimental group exposed to the combination of Prime & Bond®, LPS-PG, and Nifedipine (P<0.001). Oxidative stress was observed in experimental groups exposed to the combinations of PMMA, LPS-PG, and Cortisol (P<0.001); also noted in all three dental adhesives in combinations with LPS-PG and Nifedipine (P<0.001). Cellular membrane damage was evident in experimental groups exposed to the combinations of PMMA, LPS-PG, and Cortisol; PMMA, LPS-PG, and Nifedipine; and Prime & Bond®, LPS-PG, and Nifedipine. In conclusion, it appears that the addition of Nifedipine in combination with LPS-PG negatively alters cellular function by causing membrane damage and inducing oxidative stress.

Sustained Release of Demineralized Bone Matrix Protein in Osteoporotic Rats.

Demineralized bone matrix protein (DBM) was considered highly effective in stimulating bone healing. The objective of the study was to explore the use of tricalcium phosphate (TCP) delivery system to continuously deliver DBM in an osteoporotic condition and to evaluate changes in bone density and preservation of the spine. Ovariectomized Sprague Dawley rats were divided into three equal groups (n=16 per group). Animals in group I served as control, animals in groups II and III were surgically implanted with either empty (SHAM) or DBM filled TCP implants adjacent to L4/L5. Eight animals from each group were euthanized at 2 and 8 weeks post implantation. Femurs were evaluated for changes in density, and the lumbar spine was evaluated for changes in the endplate. Results of this study revealed (1) TCP implants were capable of delivering DBM for long duration, (2) use of sustained delivery of DBM did not induce untoward effects in the vital organs or in the uterus, fallopian tubes, or vaginal tissues, (3) DBM had no effect on chondrocyte differentiation in the spine, and (4) DBM did not increase bone density in osteoporotic female rats.

Health Impact of Retinoic Acid (ATRA) on Ovalbumin-Sensitized F344 Rat Lung and Improvement of Tissue Pathology by Citral.

The health impact of retinoic acid (All Trans Retinoic Acid; ATRA) in the development of lung pathology and tissue remodeling has not been well established in the literature. Equally, the role of Citral (inhibitor of retinoid function) in the improvement of lung pathology has not been ascertained in vivo. Therefore, it is hypothesized that ATRA and Ovalbumin (Egg albumin; OVA) exposure will sensitize lung tissues leading to lung tissue pathology and that citrals (C1 and C2) will reverse or ameliorate the related pathological damage to lung tissues. The study used an IACUC approved between-subject in vivo randomized split plot factorial design (F344 rat model; N=35). Animals were sensitized to OVA and then exposed to six different treatments; negative control (-ve), ATRA, Citrals (C1 and C2) and their triple combinations (OVA+ ATRA + C1, OVA+ ATRA + C2), by intra-peritoneal route. Rat weight data and blood were collected on Days 1 and 21, all animals were sacrificed on day 21, and lung tissues were processed for histopathology. Results from rat weights and blood (ANOVA and Duncan) as well as from the histopathological analysis of exposing the F344 rats to OVA in combinations with ATRA and citrals, revealed various levels of lung tissue damage that was impacted by exposure to citral. We conclude that OVA+ATRA+C1 combination treatment did improve lung pathology as compared to single individual treatments. However, the OVA+ATRA+C2 combination not only failed to improve these parameters, but even worsened the lung pathology of this model. This promising study showed variable responses on the interaction of Ovalbumin, citrals, and ATRA as related to their damage/improvement of related lung tissue pathologies.

The Negative Impact of Combining Retinoic Acid (ATRA) and Mold Spores on F344 Rat Lung and Improvement of Tissue Pathology by Citral.

The impact of retinoic acid (All Trans Retinoic Acid; ATRA) and Mold spores (MLD) in the development of lung pathology and in vivo tissue remodeling have not been well established in the literature. In addition, the role of citral (inhibitor of retinoid function) in the improvement of lung pathology has not been ascertained in animal studies. Therefore, it is hypothesized that ATRA and Mold (MLD) exposure will sensitize lung tissues leading to lung tissue pathology and that Citrals (C1 and C2) will reverse, ameliorate or improve the associated pathological damage to lung tissues. The study used an IACUC approved between-subject in vivo randomized split plot factorial design (F344 rat model; N=40). Animals were exposed to eight different treatments including vehicle, MLD, ATRA, Citrals (C1 and C2) and their MLD combinations (MLD+ ATRA, MLD+ C1, and MLD+ C2) by intra-peritoneal route. Rat weight and blood data were collected on Days 1 and 21, all animals were sacrificed on day 21, and lung tissues were processed for histopathology. Results from weight and blood data (ANOVA and Duncan) as well as from histopathological analyses supported the findings that exposure of F344 rats to MLD combinations with ATRA and Citrals showed various levels of lung tissue damage that were impacted by either C1 or C2. This promising study showed impressive responses on the interaction of MLD, Citrals, and ATRA as related to their impact on associated lung tissue pathologies.

The Effects of Metformin and EGCG On PANC-1 Cell Survival.

There is less than a thirty percent survival rate for patients with a localized pancreatic tumor, and less than a ten percent survival rate for patients with metastases. Patients with pancreatic cancer often have altered glucose metabolism and are prescribed metformin which has been shown to reduce cancer cell proliferation. Metformin administered at doses ranging between 10-20 mM has been reported in the literature to induce AMPK signaling pathways which increase cellular apoptosis. Epigallacto-catechan (EGCG) is a polyphenolic antioxidant that has also been shown to increase the AMPK pathway that increases cellular apoptosis. The objective of this study was to investigate the effectiveness of EGCG with a clinical dose of metformin (10µM) in reducing the survival of a pancreatic like cell line in culture. PANC-1 cells were plated onto three 24 well plates at a density of 1 x 106 cells per well. The experimental design consisted of four equal groups: Group 1 served as the control and groups 2-4 were treated with metformin, (EGCG) or metformin and EGCG, respectively. Biochemical and morphological evaluations were conducted following standard lab protocols. Results of this study show 10µM of metformin was unable to alter cell growth or proliferation over a 72 hour period, while 50µM of EGCG alone or in combination with metformin were capable of reducing cell density and cellular protein levels at 48 and 72 hours following treatment. The results show EGCG induced changes in cellular morphology which are characteristic of apoptosis. Overall, additional studies are needed to determine the effects of EGCG on AMPK and ATM pathways that are responsible in normal cellular apoptotic processes.

The Effect Of Selective Serotonin Reuptake Inhibitors and Antidiabetic Drugs on Cardiomyocytes.

Recent reports in the literature show an increase in the risk of heart related events in patients treated with tricyclic antidepressants. There is also evidence that serotonin reuptake inhibitors (SSRIs) are negatively associated with heart failure. The objective of our study is to determine if cardiomyocytes in culture can be used as a tool to mimic clinical scenarios and to evaluate therapeutic concentrations of SSRIs (fluoxetine) and antidiabetic (troglitazone) medication. Cardiomyocytes were grown in a tissue culture environment and challenged with therapeutic concentrations of SSRIs alone or a combination of SSRIs and antidiabetic drugs. Intracellular markers for stress and cytomorphometric analysis indicated SSRIs and SSRIs in combination with antidiabetic drugs negatively impact the health of the cardiomyocytes with time in culture. Analysis of the nuclear area and cytoplasmic changes are sensitive enough to use a cell-based model for determination of adverse effects associated with co-administration of drugs.

The Tissue Implant Response Surrounding Subcutaneous TCP, HA, And ALCAP Bioceramics.

The objective of this investigation was to quantify and further elucidate the tissue-implant response in the fibrous tissue surrounding tricalcium phosphate (TCP), hydroxyapatite (HA), and aluminum calcium phosphate (ALCAP) implants when implanted subcutaneously. Sixteen animals in four experimental groups (n = 4/group) were implanted with one implant each: Group I (control, TCP), Group II (HA), and Group III (ALCAP). At 90 days post-implantation, the fibrous tissue surrounding the implants was harvested. Sections of stained fibrous tissue were evaluated for the presence of macrophages, fibrocytes, neutrophils, vascularity and thickness for all three groups using semi-automated quantitative methods. The analysis indicated Group III demonstrated a significantly higher number of neutrophils but fewer macrophages and blood vessels per high power field and had a substantially thinner fibrous tissue capsule thickness compared to Groups I and II (alpha=0.05). Group II elicited a greater response of fibroblasts compared to Groups I and III suggesting HA may provide a slightly higher degree of stability to the implant. In total, these findings suggest both TCP and HA behave similarly in vivo when compared to ALCAP and may be better choices for subcutaneous soft-tissue application compared to ALCAP.