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.

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.

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.

Experimental induction of lung damage in the f344 rat upon exposure to citral, retinoic Acid (atra), ovalbumin and mold spores.

The experimental impact of retinoic acid (All Trans Retinoic Acid; ATRA), citrals, ovalbumin and mold spores in the development of lung pathology and tissue remodeling are not well established in the literature. As well, the role of these agents in lung pathology was not ascertained under an in vivo setting. Therefore, it is hypothesized that citrals, ATRA, ovalbumin and mold-spore exposure will sensitize lung tissues and will lead to the development of lung tissue pathology in these animals. The study used an IACUC approved between-subject in vivo randomized split plot factorial design (F344 rat model; N=30). Mold spores were applied to animals by intra-tracheal route whereas vehicle, ovalbumin, C1, C2 and ATRA were administered 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. Evidence from weights and blood (ANOVA and Duncan) as well as histopatholgical analysis supported the findings that exposure of these animals to C1, C2, ATRA, ovalbumin and mold spores showed different levels of lung tissue damage representing environmental exposure to these agents. This promising study showed variable lung tissue responses to the administration of ATRA, ovalbumin, citrals, and mold spores in the development of various levels of lung tissue pathologies.

Impact of paired combinations of retinoic Acid (atra) and ovalbumin on f344 rat lung tissues and improvement of related pathology by citral.

The impact of retinoic acid (All Trans Retinoic Acid; ATRA) in the development of lung pathology and tissue remodeling are not well established in the literature. As well, the role of citral (inhibitor of retinoid function) in the improvement of lung pathology was not ascertained under an in vivo setting. Therefore, it is hypothesized that ATRA and ovalbumin 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=40). Animals were exposed to 8 different treatments including vehicle, OVA, ATRA, citrals (C1 and C2) and their ovalbumin combinations (OVA+ ATRA, OVA+ C1, and OVA+ 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 weights and blood (ANOVA and Duncan) as well as from the histopatholgical analysis supported the findings that exposure of F344 rats to OVA combinations with ATRA and citrals showed various levels of lung tissue damage that was improved or worsened by either C1 or C2. 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 effects of sustained delivery of estrogen and demineralized bone matrix proteins on bone in ovariectomized female rats - biomed 2013.

Osteoporosis affects over ten million persons within the United States and is estimated to cost the healthcare system $18 billion dollars a year. Approximately 1.5 million persons will be diagnosed with an osteoporotic fracture and the epidemiological data reflects that prevalence of the osteoporotic fractures is four times more common than having a stroke. The current treatments strategies for osteoporosis are geared toward inhibiting the osteoclast cell resorption of bone, and not on the osteoblast bone formation. The use of demineralized bone matrix proteins (DBM) has been shown to be effective in healing osteoporotic bone fractures within a four week time period. Our goal was to deliver in a sustained manner DBM over an eight week period and compare bone strength and bone histology to osteoporotic untreated animals, osteoporotic animals given sustained delivery of physiological estrogen, as well as naïve control (animals with ovary intact). Our results showed estrogen administered in a sustained fashion was able to reverse the decline in bone strength and re-establish the bone quality similar to ovary intact controls. DBM administered in a sustained manner showed similar bone quality and strength to osteoporotic control animals. Administration of DBM to mature bone in a sustained fashion may be ineffective in inducing osteoblast function or reversing osteoclast activity. It is possible that DBM may be more effective on immature bone cells.

The effects of sustained delivery of corticosterone on the adrenal gland of male and female rats - biomed 2013.

Glucocorticoids have long been recognized to have beneficial effects in rheumatoid arthritis and asthma. Numerous clinical trials show the efficacy of short term low dose treatment to resolve inflammation. Despite the success of short term use, there is concern regarding chronic use of glucocorticoids because of the development of exogenous Cushing’s syndrome. Chronic variable stress models have detailed the effects of chronic stress exposure on body weight, plasma corticosteroid levels, ACTH levels, and adrenal weights, but limited studies detail the effects of the body systems induced by continuous exposure to glucocorticoids similar to that seen in exogenous Cushing’s syndrome. The present study uses a TCPL drug delivery system to administer corticosterone (CS) continuously in male and female animals for 24 days and evaluates long term chronic use effects on body weight, adrenal weight, and adrenal ultrastructure. Continuous release of CS resulted in slight decreases in body weight in both male and female rats and decreases in adrenal wet weight in the female rats. Ultrastructural changes were seen in the adrenal histology in both female and male rats. Male rat adrenal glands showed atrophy of the zona glomerulosa and hypertrophy of the adrenal medulla. Female rats showed disorganization of all zones within the adrenal gland and an increase in fat around the gland. The information is important for understanding physiological differences in males and females during stress. The continuous release of CS may provide insight into the pathology of exogenous Cushing’s syndrome.

Reproductive system behavior following exposure of sustained delivery of npy antagonist in ovariectomized (ovx) rats.

Several investigations have documented that sustained delivery of estrogen can modulate or sustain normal female reproductive functions. However, the literature is lacking scientific evidence regarding the mechanism of estrogen and neuropeptide Y antagonist (NPY) effect on the hypothalamic-pituitary-gonadal axis. The objective of this study was to explore the role of sustained delivery of estrogen and its effects on reproductive unction compared to an antagonist such as NPY. A total of twenty adult female rats (OVX, n=15; intact control, n=5) were divided into five groups (intact control, OVX, sham, OVX + estrogen, and OVX + NPY). Animals in two groups were surgically implanted with a TCP delivery device loaded with estrogen or NPY. Vaginal smears and body weights (BW) were evaluated at baseline and at two weeks post implantation. At the end of two weeks, all animals were euthanized and vital and reproductive organs were retrieved for histopathological evaluation. The results revealed differences in BW between intact control and OVX animals. Furthermore, there was statistical difference (P<0.05) in BW between OVX and OVX + NPY animals. Vaginal smear evaluation revealed that estrogen exposure induced estrus cyclic activities as compared to OVX and sham animals. The animals exposed to sustained delivery of NPY triggered moderate cyclic activities compared to intact control animals. There were no significant differences (P<0.5) in vital organ wet weights among and between animals in all groups. Overall this study proved the capability of TCP to release estrogen and NPY at sustained levels, which resulted inpathophysiological changes in female reproductive organs.

Evaluation of the estrous cycle and ovarian function following sustained delivery of corticosteroid via tcpl delivery - biomed 2011.

Stress has been shown to impair reproduction in many species and the hypothalamic pituitary axis (HPA) has been shown to be the target. Most of the literature evaluated the effects of acute stress and focused solely on the HPA. The literature is lacking in the area of chronic administration of corticosteroid use and effects on the estrous cycle and long term effects on the ovary. The objectives of this study were to use tricalcium phosphate drug delivery systems to deliver corticosterone in a sustained supraphysiological level and follow the estrous cycle over a 28 day period; then, harvest the ovaries to evaluate the morphology. The results indicate prolonged administration of corticosterone in female rats cause alteration in the cyclic activity as evidenced by changes in the estrous as well as morphology of the ovaries. The mechanism appears to be disruption or interference of the HPA axis. Overall, ceramic drug delivery devices can be used effectively to deliver corticosteroid to induce pathophysiological changes associated with stress.

The effect of agonists and antagonists on Hep-2 cells.

Androgens may play an important role in promoting the growth of laryngeal carcinomas. The aims of this investigation were to investigate the effects of (testosterone (T) and androstendione (AED)) in the presence of the anti-androgen, spironolactone (S), n Hep-2 cellular proliferation and damage after 24, 48 and 72 hours. Hep-2 cells were divided into six groups (n = 5) control, S, T, AED, S+T, and S+AED, respectively. The cells were harvested after each incubation period into two different fractions: suspended cells and adhered cells. Cell counts and cellular damage determinations were performed on each fraction. Analysis of variance was used to determine significance at p < 0.05. Data for cell counts revealed an interesting phenomenon between the two fractions. Adhered cells showed decreased cell numbers in the presence of S and T for 24 - 48 hours followed by a significant increase at 72 hours. Cells in the adhered fraction incubated in the presence of AED or AED + S were significantly lower for the duration of the experiment. However AED or AED + S treatment caused significant increase in cell number in suspended fraction for the duration of the experiment. All treatments after 72 hours showed a slight reduction in MDA levels indicating treatments did not cause cell damage. Overall, the data suggests the possibility of two populations of cells that respond differently to the AED. T had no significant effect on either cell fraction for the first 48 hours followed by a significant increase in cell number at 72 hours suggesting T may need to be converted enzymatically to the more potent androgen, dihydrotestosterone.

The effects of genistein concentrations on Hep-2 cellular function.

Genistein is a phytoestrogen that has shown potential as a chemotherapeutic agent, which acts by inhibiting protein-tyrosine kinase and topoisomerase II enzymes. These particular enzymes are crucial for cellular proliferation. The goal of this study was to evaluate the effect of genistein concentration (0.5, 0.05 or 0.005 mg/mL) on Hep-2 cells functional capacity. Specifically, to evaluate cellular number, protein, damage and morphology at 24, 48, and 72 hours phases. Data obtained from this study revealed that cell numbers were significantly reduced in low and medium concentrations after 24hrs, and cell numbers appeared to rebound at 48 and 72hrs in an inverse fashion. This data suggests continuous administration of the drug at therapeutic levels would serve as a better chemotherapeutic agent. Cellular damage was not evidenced and suggesting that the drug did not target the cellular membrane site. Morphological changes such as anucleation were seen at 24 hrs in all doses suggesting that genistein targets the genome directly. Interestingly, cellular function was able to recover in the lower doses of genistein treatment indicating cellular metabolism of the drug. Also, this information suggests that genistein mode of action by targeting enzymatic activity, as opposed to causing alterations within the cellular membrane, leading to leakage of cellular contents and ultimately cellular death since the membrane did not show evidence of lipid peroxidation.

Growth and cell viability of estradiol and IP-6 treated Hep-2 laryngeal carcinoma cells.

Inositol 6-phosphate (IP-6) has demonstrated novel anti-cancer activity using several different tumor models. IP-6, a phytoestrogen, has estrogen receptor (ER) binding capabilities that are not known to cause cellular proliferation in hormone sensitive cells. It is hypothesized that IP6 can induce competitive inhibition with estrogen for estrogenic binding sites on cancer cells resulting in decreased proliferation. In this experiment, Hep-2 cells were treated with Estradiol and IP-6 in a dose dependant manner for 24, 48, and 72 hours. They were analyzed for changes in number, protein concentrations, damage, and morphology. There was an increase in cell proliferation in Estradiol treated Hep-2 cells. Cells treated with IP-6 showed no change in cell proliferation in the 24 or 48-hour groups, but there was a decrease in number with the 72-hour group, particularly with the 1mM dose. Both the Estradiol and IP-6 treatments caused no membrane oxidation and the level of protein synthesis stayed consistent. The morphology showed small round to cuboidal, single cells with scant, dense, basophilic cytoplasm and hyperchromatic nuclei with smooth borders. Some cells showed anucleation and cellular degeneration. Although IP-6 is a phytoestrogen, the results show that affinity for estrogen binding sites on Hep-2 cells is greatly decreased. However, with time given increased concentrations, IP-6 can cause a decrease in the cellular proliferation of Hep-2 cells without initiating cellular apoptosis.

The effects of isolated antioxidants from black seed on the cellular metabolism of A549 cells.

UNLABELLED: Reactive oxygen species cause cytotoxic effects. The body requires the uptake of exogenous compounds with antioxidant potential. Black seed (BS) is a plant that contains at least one active lipid soluble antioxidant, thymoquinone. Fractionation of the seed components yielded antioxidant compounds in both the water soluble and lipid soluble fractions. The objective of the study was to determine the safety of the fractionated compounds and compare their potency with pure thymoquinone and vitamin E on A549 cells in culture for 24, 48 and 72 hours. RESULTS: Black seed extracts and pure thymoquinone showed markedly reduced levels of MDA for the duration of the study. The vitamin E dosage used led to greater toxicity and cellular damage rather than cell protection. Cellular proteins levels after 24 hours showed cells in BS + ETOH extract group had the highest metabolic activity. However, at 72 hours, the activity was shifted and showed the least amount of protein synthesis. As for vitamin E, the results were consistent throughout all three phases showing slow metabolic activity. Cell number was decreased after 24 hours in thymoquinone treated cells, and remained reduced for the duration of the study. The BS+H2O fraction showed a similar trend to thymoquinone, where as the BS+ETOH fraction showed a negative shift in cell number at 48 hours when compared with the control. CONCLUSION: The study concludes that of the two BS extracts, the BS+H2O extract had the greatest effect on the cell viability and function.

Delayed apoptosis upon the treatment of Hep-2 cells with black seed.

Nigella sativa (Black seed, BS) has been used to promote health and fight disease for centuries. The objectives of this investigation were: (1) to study whether agents such as cortisol and LPS alone or in combination induce cellular (Hep-2, laryngeal carcinoma) damage with time in culture (24, 48, and 72 hours) using apoptosis as a marker, (2) to determine if an immune stimulant such as BS, can protect Hep-2 cells from insult and ultimately thwart the programmed cells death mechanism. A total of 54 Hep-2 cell/tubes (50,000 cells per tube) were divided into six equal groups. Group one served as untreated control, while groups 2-6 were treated with either cortisol (10 ng/ml), LPS (10 micrograms/ml), BS (25 micrograms/ml), or a combination of LPS and cortisol and cortisol plus LPS plus BS, respectively. At the end of each phase the cells were harvested, heat fixed and stained with H&E to evaluate morphological changes. Immunohistochemistry, using antibodies against caspace-3 to evaluate cells undergoing apoptosis was conducted in all groups. The results of this study showed evidence of cells undergoing apoptosis at different magnitudes in all groups. However, the most dramatic change was seen in groups containing cortisol and LPS alone or in combination. This was supported by the fact that there were several adaptive responses observed in all phases. In addition, the exposure of BS to cells pretreated with cortisol and LPS showed evidence of protection against the progressive apoptosis.

The role of black seed in the proliferation and biochemical marker levels of Hep-2 cells.

For centuries, people in the Middle East and Southeast Asia have used Nigella sativa, also known as black seed (BS), for its homeopathic effects. The objective of this study was to investigate the role BS might have on the metabolic biomarkers of the Hep-2 cell line. The experimental design entailed six groups of five wells each (50,000 cells). Groups II through VI were treated with BS, lipopolysaccharide (LPS), cortisol, LPS + cortisol, and BS + LPS + cortisol, respectively. Group I was the untreated control group. At the end of 24, 48, and 72 hours, the total cell count, protein and MDA levels were measured by following standard lab protocols. Data collected from this study revealed that Hep-2 cells exposed to LPS and cortisol (group V) resulted in a decrease in cell proliferation compared to the control. BS treatment induced a higher proliferation rate than group V. Similar trends were observed in the metabolic behavior of Hep-2 cells as evidenced by the total protein and MDA levels. The exposure of BS showed a shift in the metabolic pathways. In conclusion, this study showed that exposure to LPS resulted in an alteration in the metabolic function and this phenomenon was further escalated under stressful conditions (increased cortisol exposure). In addition, the use of BS reversed the traumatic condition.

Morphometric evaluation of ovarian tissue exposed to PCB conventionally and in a sustained manner.

Most natural estrogens are short-lived, do not accumulate in tissue and are easily broken down in the liver. In contrast to natural estrogens, estrogenic drugs such as ethynylestradiol, diethylstilbestrol, synthetic environmental estrogens such as beta-hexachlorocyclohexane, polychlorinated biphenyls (PCBs), and phytoestrogens, are more stable and remain in the body longer than natural estrogens. Because most of these compounds are lipophilic, they tend to accumulate within the fat and tissue of animals and humans. Thus, depending on the natural estrogen levels, environmental estrogens may have different influences (mimicking, or blocking estrogen's effects) on estrogen activities. A total of 14 adult female rats were divided randomly into three groups. Animals in group I (n = 4) served as control. Animals in group II (n = 5) were injected with PCBs, and animals in group III (n = 5) were implanted with TCPL capsules containing PCBs. Pap smears were obtained daily, and at the end of the experimental phase the animals were sacrificed and vital organs as well as reproductive organs were obtained and wet weights were recorded. Significant reductions in ovarian wet weights were found in all animals treated with either sustained release of PCBs or injection of PCBs. Ovarian tissue was further analyzed histologically to determine the effects of PCBs. Histomorphometric data revealed significant reduction (p < 0.05) in the total ovarian area of animals treated with PCBs. Measurements of cross-sectional lengths confirmed the reduction seen in the area. Overall, the data suggest that PCB pollutants have produced detrimental effects on endocrine function as well as fertility regardless of the route of administration.

Morphological evaluation of MRC-5 fibroblasts after stimulation with static magnetic field and pulsating electromagnetic field.

The quality of tissue repair and the speed with which that repair can be accomplished are the two major variables in the healing of any injury. Today, magnetic field exposure to traumatized areas has shown to be a promising tool in the healing process. The exact mode of action by which radiating and unchanging magnetic fields still has to be elucidated. The objective of this study was to evaluate the morphology of MRC-5 fibro-blasts after stimulation with static and pulsating magnetic fields. Under sterile environment, a total of 24 wells were loaded with 50,000 MRC-5 cells each and further divided into three groups. Groups 1 and 2 were exposed to magnetic fields, static and pulsating respectively. Group 3 wells were unexposed and served as the control group. The cells were monitored at 0, 24, 48, and 72 hours and representative views were captured using digital analysis techniques. The live cells were screened for cellular mobility, cell distribution, and cellular morphology (size, shape, lysis, and background). After 72 hours, the supernatants and cells of all three groups were collected and MDA analysis was performed to determine possible cellular damage. Group 1 cells continued to grow at a reasonable rate, but there was substantial cell membrane damage (high MDA levels, p < 0.05). Group 2 cells appeared to be very stressed under these conditions especially at the initial phase (24 hours). In conclusion, the use of pulsating magnetic stimulation can be beneficial in the healing process of soft tissues.

Down regulation of CD14 expression through pretreatment with glucocorticoids.

Glucocorticoids such as cortisol are potent immunosuppressive agents that act on many cells of the body, including macrophages. Macrophages express CD14 in response to lipopolysaccharides (LPS) extracted from bacterial coats. The objectives of this study were: (1) to determine if pretreatment of macrophages with cortisol for 30 minutes prior to challenging the cells with endotoxin results in increased cell loss, cell damage (MDA), and suppression of CD14 receptors; and (2) to determine if CD14 receptor expression is able to recover with time. An experimental design incorporating RAW 264.7 cells (RAW) was used in order to evaluate our objectives. The cells were plated on 24 well plates and subsequently divided into four groups. The first group was untreated and served as the control. Group two was treated with LPS, group three with 10 uL of cortisol and a combination of LPS and cortisol was used in the treatment of the fourth group. The cells were recovered at 24, 48, and 72 hours. Results showed that there was a significant decrease in the proliferation rate in RAW cells exposed to cortisol and LPS either alone of in combination when compared to the untreated cells. Cell damage was also increased in treated cells. LPS caused receptor expression at all time points. CD14 expression was down regulated at 48 hours in cells pretreated with cortisol, however, this suppression was no longer evident at 72 hours.