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This study investigates conventional delivery of thymoquinone (TQ) and epigallocatechin-3-gallate (EGCG) when applied to Caov-3 and SK-OV-3 ovarian like cell lines. Culturing the cells and assessment of functional activities were conducted following standard lab protocols. The results revealed: (1) an increase in the SK-OV-3 cell protein levels following treatment with TQ+EGCG which was statistically different (p

It is well documented that glucocorticoids are potent anti-inflammatory and immunosuppressive agents that are known to affect cell mediated inflammation by the inhibition of cellular proliferation and cytokine production. The literature is lacking knowledge in elucidating the mode of action of such agents on the transformed like cells in culture. Therefore, RAW (macrophage like) cells was selected as a model to determine the effects of cortisol administration or cortisol in the presence of LPS on the cells metabolic functions. Cells were treated with physiological concentrations of cortisol or cortisol + LPS for periods of 24, 48 and 72 hours. After each phase, cell numbers, cellular damage and cellular morphology were determined. The results indicated cortisol and cortisol + LPS treated cells inhibited cellular proliferation as well as increased cellular MDA levels as early as 24 hours. Overall, the results indicate that cortisol has a remarkable effect on RAW cellular proliferation similar to the reduction seen in our previous findings using HEP-2 cells. In addition to reduction in cellular number the cell's ability to adjust to a bacterial challenge may be directly altered. These results provided important information for patients who are immunosuppressed or chronically exposed to stressful conditions.

The route of administration of ovarian hormones and other biologicals represent a key factor in the treatment, prevention or eradication of different diseases in humans. Many studies have documented that the use of sustained delivery of biologicals is more effective compared to conventional route of administration. To date, the literature is lacking studies that conclusively demonstrate the change on body weights and vital organ morphology associated with sustained delivery of Neuropeptide Y antagonist and estrogen compared to intact and ovariectomized adults as a model. Therefore, the purpose of this study represents a histopathologic evaluation of the sustained delivery of ovarian hormones and neuropeptide y antagonist on the body weight and vital organs of ovariectomized rats. A total of 25 Sprague Dawley rats were obtained and divided into five groups, intact control with ovaries, sham, OVX + estrogen, and NPY antagonist. Animals in three of the five groups were surgically implanted with a TCP drug delivery device loaded with 1.6 mg of estrogen and 1.6 mg of NPY antagonist, and the sham animals were implanted with empty capsules. The capsules were surgically inserted into the muscle adjacent to the six lumbar vertebra. The animals were euthanized at two weeks, post-implantation phases. An analysis of the body weights were compared, and the OVX + estrogen and the OVX + NPY antagonist were similar to the intact control animals. Histopathologic evaluation of the major body organs revealed no histopathological change in the adrenals, spleen, liver, lungs or heart muscles of the intact, OVX, sham and NPY antagonist low and high dose treated animals. The only significant difference was observed in the kidney, where the glomeruli appeared much larger in the estrogen treated animals. Overall conclusion obtained from this study demonstrated that TCPL delivery can be used effectively to administer NPY at sustained level with remarkable reduction in side effects that associated with conventional means.

The purpose of this investigation was to determine fibroblast behavior after implantation of ultra-high molecular weight polyethylene (UHMW-PE) rinsed with saline (control) or coated with poly-L-lysine (PLL), arginine-glycine-aspartic acid (RGD), or arginine-glycine-glutamic acid (RGE) into 16 adult male rats subcutaneously. At 90 days post-implantation, fibroblast counts were highest in the saline rinsed group (34±2 cells/HPF) and significantly reduced in RGD (19±10), RGE (2±3), and PLL (0) treated groups. These findings indicate fibroblast migration in surrounding fibrous tissue can be strongly influenced using various amino acid combination coatings in subcutaneous applications.

The objective of this study was to investigate the effect of temperature of sintering of tricalcium phosphate ceramic (TCP) on the adherence and viability of monocyte and monocyte derived macrophages. The monocytes were isolated from human peripheral blood, biotin labeled and seeded at a density of 5/spl times/10/sup 5/ cells/well according to standard laboratory procedures. Cells were considered macrophages after remaining in culture for 24 hours. Cells were then plated in each microtiter well preloaded with TCP of different sintering temperatures (400, 800 and 1150/spl deg/C). At the end of 1, 2, 3 and 7 days the viability and cell number of monocyte or monocyte derived macrophages were determined using an established assay (biotin-labeled-macrophages). Cell number was determined in control wells with known amounts of cell number, a standard curve was generated by plotting absorbance units versus cell number. At the end of each phase, the blocks were collected and prepared for SEM evaluation. The data from this experiment suggest that: (i) monocytes and macrophages are capable of surviving on all different types of blocks tested over a 7 day period, (ii) SEM evaluation of the ceramic blocks showed an increase in the size and number of micro and macropores with time in blocks sintered at 400 and 800/spl deg/C. In contrast, the size and number of pores calculated from representative views of TCP blocks sintered at 1150/spl deg/C were significantly lower than the other two groups. The mechanism of action of these cells toward the aforementioned bioceramic surfaces has to be investigated in a larger scale. Information obtained from this study provided new insights on the interrelationship between bioceramics, temperature of sintering effect, incubation time and the possible cell response during chronic inflammation at the site of implantation.

Tissues retrieved during revision arthroplasty (n=25) were processed for routine immunohistochemical techniques to localize the presence of bone resorbing cytokines (IL-1 and IL-6), and to identify areas of significant necrosis and metallic deposition. Tissues were processed and embedded according to standard laboratory techniques. Several slides were prepared for each tissue sample taken. Several sequential slides were stained for Hematoxylin and Eosin to demonstrate cell type. Other slides of the same tissues were processed for localization of cytokines, IL-1 and IL-6. To identify specific cytokine reactions, the tissues were first blocked with proteins to ensure that all. The results revealed that there were several locations on the tissues that appeared highly reactive for cytokines IL-1 and IL-6. However, some of these areas turned out to be nonspecific when compared with negative controls and the Hematoxylin and Eosin stained slides. Areas of extreme necrosis or where metal debris was presence demonstrated sites of nonspecific reactivity. Areas of necrosis have a high nonspecific affinity for antibodies and these areas of necrosis can be demonstrated on the Hematoxylin and Eosin sections, and should be avoided in determining the amount of specific cellular reactivity. In addition, new immunohistochemical techniques use cobalt ions as enhancers of peroxidase response, in areas on slides labeled as negative controls where metallic debris was trapped, there was an increase in nonspecific positive reactivity. These factors must be carefully controlled in determining true cellular responses that causes resorption of bone and implant failure.

The relationship between various bioceramics (similar to bone) used in surgical implantation and osteoclastic response has not been fully elucidated. The objective of this study was to investigate the effect of various biomedical ceramics such as tricalcium phosphate (TCP), hydroxyapatite (HA), and aluminum-calcium-phosphorus oxide (ALCAP) on the adherence and viability of mice osteoclast (OT) in vitro. The OT cells were isolated from adult male mice and seeded at a density of 5/spl times/10/sup 5/ cells/well according to standard laboratory procedures. Cells were plated in each micrometer-well pretreated with ceramic capsules (HA, TCP and ALCAP) and buffered control. At the end of 1, 2, 3 and 5 days the viability and cell number of OT were determined using an established assay. Cell number was determined in control wells with known number of cells, and a standard curve was generated by plotting absorbency units versus cell number. Biochemical analysis was performed on the aliquots obtained from the experimental and control wells at the end of each phase of the investigation. The data from this experiment suggest that: (i) OT are capable of adhering to the surface of HA, TCP and ALCAP in an in vitro environment for over a 5 day period; (ii) Long term incubation of ceramic capsules with OT revealed that the cells experienced gradual disassociation phenomenon with a greater numbers of cell detachment seen in the ALCAP contained wells; (iii) SEM analysis of representative capsules demonstrated that there is an increase in the number of micropores and macropores on the surface of the materials after contacting a cellular environment. This observation suggest that the material surface has been modified (TCP>HA=ALCAP). Information obtained from this study provided new insights on the interrelationship between bioceramics and the possible OT response during chronic inflammation at the site of implantation.

It is well documented that testosterone stimulates protein anabolism, muscular development, bone growth and increased metabolic rate. In addition, exogenous low level of testosterone can inhibit the secretion of gonadotropins by the anterior pituitary. The objective of this study was to investigate the effect of sustained delivery of testosterone enanthate (TE) on the testicular architecture by means of tricalcium phosphate lysine (TCPL). In this experiment adult male Sprague Dawley rats (250-300 g BW) were randomly divided into three equal groups (n=16). Rats in group I were implanted subcutaneously with TE loaded TCP implants (4-6 ng/ml). Rats in group II were implanted with sham TCP capsules, and rats in group III served as intact unimplanted controls. Surgical aseptic techniques were performed according to standard laboratory procedures. At the end of 2 and 4 weeks post implantation, 8 animals from each group were sacrificed and the testes were collected, weighted, and embedded for histopathological evaluations. The results of this study revealed the following: (i) Sustained delivery of TE reduced spermatogonia numbers in animals after 2 and 4 weeks by approximately 10% in comparison to control animals. (ii) The lumen of the tubules showed slight regression after 4 weeks of TE treatment. (iii) Overall decrease of the vasculature of the experimental animals was seen after 2 and continued through 4 weeks with an occasional increase of microvasculature seen in animals treated for 4 weeks with TE. (iv) Finally, the germinal epithelium showed significant reduction in secondary spermatocytes and conversion to mature sperm.

Fibrous tissue formation is often used as a screening method to determine biocompatibility of orthopaedic and dental implanted material. In this investigation, porous implants of hydroxyapatitelysine (HAL) delivery devices filled with Vitamin E were implanted intraperitoneally (IP) using adult male rats as a model. The fibrous tissue surrounding the implant was studied histochemically to determine the resorbability rate of the devices. Fibrous capsular tissues were carefully dissected away from the capsule noting the tissue closest to the implanted material. Evaluation of the sections (5 /spl mu/m, H&E) collected from various areas of the implants (n=5 per group) revealed that: (1) all Vitamin E-HAL capsules were encapsulated with fibrous tissue within seven days post-implantation and the thickness of the fibrous tissue was found to be directly proportional to duration of implantation, (2) the thickness of fibrous tissues collected from IP site were significantly higher (22%/spl plusmn/3.4 SD) than those collected from SC site of implantation in another study, (3) vascularity, macrophages, fibroblasts, and plasma cells at the Vitamin E-HAL implant-interface were evident after the first week, and the persistence of these cell types and progressive angiogenesis in subsequent weeks was apparent. The formation of distinct cell layers was remarkable in the ninth week phase, (4) SEM and radiographs analysis of retrieved devices showed a significant (P

The objective of this study was to evaluate the effect of various dosages of testosterone (T) delivered in a sustained manner by means of a tricalcium phosphate-lysine (TCPL) delivery system on morphological changes of prostatic tissue using adult male rats as a model. In this experiment, adult male rats (250-300 g BW) were randomly divided into five equal groups (n=8). Rats in group I, II, and III were implanted subcutaneously with TCPL loaded with three different dosages (10, 100 and 200 mg, respectively) of T. Rats in group IV were implanted with sham TCPL capsules, and rats in group V served as intact unimplanted controls. At the end of 8 weeks post implantation, all animals were sacrificed and the prostate tissues were collected, weighted, and embedded for histo-pathological evaluations. Data collected from this study have shown that exogenous intake of T delivered in a sustained manner for eight weeks induced several pathophysiological conditions in ventral prostatic tissue in comparison to the control and sham operated groups. This phenomenon was found to be directly proportional to the dose or the level of sustained delivery. The results demonstrated that the use of 10 mg filled TCPL implants decreased the total mass weight of ventral prostate. Light microscopic evaluation of this group (Group I) revealed a cellular adaptation through an atrophy in the epithelium component. Cytopathological observations such as low cuboidal and thin glands, pleomorphism, and occasional presence of connective tissue stroma were detected. In contrast, ventral prostate collected from animals implanted with TCPL filled with 200 mg T (Group III) showed a significant increase in weights of the wet prostatic tissues in comparison to all groups. Histopathological evaluations demonstrated the following: (i) prostatic hypertrophy alone, or in conjunction with hyperplasia of the epithelial cells, (ii) less connective tissue stroma in comparison to the control group, (iii) occasional involvement of mitotic figures, and (iv) increased angiogenesis. No significant change was observed in those animals implanted with TCPL capsules containing 100 mg T compared to the intact control animals.