Gelatin Carrier Research Articles

Short-term study of retinal pigment epithelium sheet transplants onto Bruch's membrane

The purpose of this study is to investigate the survival and behaviour of retinal pigment epithelium sheets transplanted onto hydraulically debrided Bruch's membrane. Uncultured retinal pigment epithelium sheets obtained from male cats and sandwiched between two gelatin sheets were transplanted onto the tapetal area of female cats after native retinal pigment epithelium was debrided. For controls, the gelatin carrier was transplanted after debridement. Each transplant or control specimen was analyzed histologically and immunohistochemically. Transplanted male retinal pigment epithelial cells were identified by in situ labelling of the cat Y chromosome. Over half of the transplants appeared as retinal pigment epithelium multilayers in the subretinal space. Retinal pigment epithelium pigment dispersion into the subretinal space was seen in most of the transplants, and retinal pigment epithelium pigment infiltration into the neural retina was seen in all 7-day survival transplants. A few condensed darkly stained retinal pigment epithelium nuclei and Terminal Transferase dUTP Nick End Labelling-positive retinal pigment epithelium cells were observed in all transplants. Cellular retinaldehyde-binding protein was present up to day-7 in most transplanted RPE cells. In both transplant and control specimens, the antibody against the Ki-67 nuclear antigen labelled a few retinal pigment epithelium cells at day-3. Terminal Transferase dUTP Nick End Labelling-positive outer nuclear layer nuclei were most frequently observed at day-1 but were much less frequent at day-3 in both transplants and controls. The survival and effectiveness of retinal pigment epithelium sheet transplants appeared similar to the retinal pigment epithelium microaggregates transplants conducted previously in this model.

Effect of route of administration and carrier on bioavailability and kinetics of astaxanthin in Atlantic salmon Salmo salar L.

This study examined astaxanthin bioavailability and kinetics in adult Atlantic salmon Salmo salar L., following two different routes of astaxanthin administration (oral vs. intraperitoneal (i.p.) injection) using two different carriers of the pigment (gelatin vs. sesame oil). The dorsal aorta of adult Atlantic salmon (mean initial weight 950 g) was cannulated. The fish received a single dose of astaxanthin (572 μg kg−1) in sesame oil or (514 μg kg−1) in gelatin via the oral or i.p. route. Plasma was sampled regularly up to 72 h post oral administration and up to 510 h post i.p. injection. The astaxanthin concentration–time curves from plasma were best fit to a one-compartment pharmacokinetic model for each of the four treatments. The gelatin carrier resulted in higher availability of astaxanthin compared to the sesame oil carrier. The bioavailability for astaxanthin in sesame oil was only 38.7% of that in gelatin by i.p. injection, and only 53.5% of that in gelatin by oral administration. Higher availability of astaxanthin was observed when i.p. injection was used compared to oral administration. The bioavailability for astaxanthin administered orally was only 12% of that by i.p. injection in sesame oil, and only 8.7% of that by i.p. injection in gelatin.

β-Galactosidase from Kluyveromyces lactis cell disruption and enzyme immobilization using a cellulose–gelatin carrier system

Whole cell immobilization is one of the major immobilization methods due to cost advantages. Extracellular enzyme producing cells can be immobilized directly, but intracellular enzyme producing cells should be treated first to increase cell permeability. In this work β-galactosidase-producing Kluyveromyces lactis (ATCC 8583) cells were used. Since β-galactosidase is an intracellular enzyme permeabilized dead cells were immobilized into gelatin using glutaraldehyde as cross-linker. Two chemical and one physical disruption processes were tested and the physical method was determined to be better because of the probable risk of chemical toxicity accompanied with the chemical methods. Thirty percent activity was obtained by immobilized cells relative to free disrupted cells.

Leukemia inhibitory factor enhances bone formation in calvarial bone defect.

For bone defect reconstruction, locally administered cytokine plasmid was examined. Leukemia inhibitory factor (LIF) can bind to the osteoblast cell surface and induce bone formation both in vitro and in vivo. The authors investigated the local mouse LIF complementary deoxyribonucleic acid (cDNA) plasmid in the pcDNA 3 expression vector, which is promoted by cytomegalovirus and is stabilized by bovine growth hormone polyadenylation, with a gelatin sponge carrier. A total of 150 male Wistar rats were used. They were divided into three groups. Group 1 (N = 30) was treated with the gelatin carrier of the pcDNA 3 vector, group 2 (N = 90) was treated with three different doses of LIF cDNA (0.1, 1, and 10 micrograms) in the pcDNA 3-vector plasmid along with the gelatin carrier, and group 3 (N = 30) was treated with recombinant human bone morphogenetic protein -2. Ten animals in each group were euthanized at 1, 3, and 5 weeks postoperatively. Animals treated with LIF cDNA showed significantly enhanced bone mineral density (p < 0.05), as confirmed by dual-energy X-ray absorptiometry (DEXA), in 3 weeks compared with the control vehicle. By 3 weeks, the number of fibroblast-like cells and collagen fibers decreased, whereas the osteoblast-like cells increased inversely, as revealed during histological examination. LIF messenger ribonucleic acid demonstrated by in situ hybridization was observed most markedly in osteocytes of the LIF cDNA-treated group. Also, LIF peptide was detected in the same cell type by immunohistochemistry. Locally administered LIF cDNA plasmid in a gelatin carrier can increase bone density significantly, with subsequent bone formation, probably via osteocyte activation.

Preparation and characterizations of self-assembled PEGylated gelatin nanoparticles

The PEGylated gelatin nanoparticles were prepared by self-assembling method and characterized. The gelatin drug carrier was proposed as a targeting drug delivery system with the hypothesis that the gelatin carrier could be degraded by the matrix metalloprotease (MMP) and release the anticancer drug loaded inside carriers around the cancer site. The gelatin nanoparticles proposed in this study were composed of deoxycholic acid (DOCA), monomethoxy polyethylene glycol (MPEG), and gelatin. The carboxyl groups of DOCA and carboxylated MPEG were coupled with amine group of gelatin by dichlorohexylcarbodiimide (DCC) method. One molecule of gelatin coupled with 205 molecules of MPEG and 275 molecules of DOCA. The synthesized gelatin/DOCA/MPEG conjugates (GDM) were ultrasonicated to produce self-assembled nanoparticles. DOCA acted as the hydrophobic core, thereby aggregating gelatin molecules and hydrophilic MPEG chains located at the surface of the nanoparticles. The concentration of GDM, intensity of sonication, sonication time and temperature, all affected to control the particle size in the ultrasonication. The optimum condition was obtained as 1.0 mg/mL of GDM, 28 W for sonication intensity, 3 min of sonication time, and room temperature. The size distribution of particle was found to be 100–1000 nm in this condition. The particles which had a broad size distribution were filtered by 0.2 μm membrane. The product yield of particles having below 200 nm of size was about 30%. After filtration, an average diameter of GDM nanoparticle was 176 nm (155–200 nm).

Investigations on Carrier Systems for Alpha-Amylase Enzyme Immobilization

Abstract α-Amylase enzyme was immobilized onto gelatin carrier system. Chromium (III) acetate and potassium chromium (III) sulfate were used as crosslinking agents. Effect of carrier system composition on immobilization was investigated by combining different proportions of carboxymethylcellulose or polyacrylamide with gelatin. Considerable relative activity enhancements were achieved (41% for carboxymethylcellulose-gelatin and 25% for polyacrylamide-gelatin). Reusability's of immobilized enzymes were also investigated. Activitie were stable at least for 20 uses.

Immobilisation of β‐galactosidase onto gelatin by glutaraldehyde and chromium(III) acetate

Abstractβ‐Galactosidase (EC 3.2.1.23) was immobilised onto a gelatin carrier system by crosslinking. Glutaraldehyde and chromium(III) acetate were used as crosslinking agents. The effect of pH on enzyme activity, kinetic parameters and reusability of the immobilised enzyme were investigated. To obtain a better relative activity, a method was developed and further modified to increase the surface area of the support system by using polyacrylamide. Relative activity enhancement of 68% was achieved in the gelatin–glutaraldehyde system.

Ethyl m-aminobenzoate methanesulfonate dependent and carrier dependent pharmacokinetics of extremely lipophilic compounds in rainbow trout

Significant differences were found in both uptake and pharmacokinetics in fish when six lipophilic compounds were dosed by gavage in either an oil or an gelatin carrier. Pharmacokinetics were also different when fish were anaesthetized with ethyl m-aminobenzoate methanesulfonate (MS-222) before dosing. The highest uptake percentages, uptake rates and concentrations of the compounds were found in the fish which were given the gelatin carrier only. MS-222 decreased the uptake of the compounds. Absorption of the compounds from oil was lower than from gelatin. In addition, absorption from oil continued for 21 d, which lasted longer than from gelatin. In the fish which were given the gelatin, the concentrations of the compounds in both muscle and liver showed a compound dependent decrease in the course of the experiment. In general, in all treatments higher concentrations were found in liver than in muscle. For each treatment, the absorption rates of pentachlorobenzene, hexachlorobenzene, 2,2′,5,5′-tetrachlorobiphenyl and 2,2′,4,4′,5,5′-hexachlorobiphenyl were similar, but different from the other treatments. It can be concluded that the experimental methodology significantly affects the pharmacokinetic parameters that are used to model the bioconcentration of hydrophobic chemicals in the environment.

Anaphylaxis to measles, mumps, and rubella vaccine mediated by IgE to gelatin

Background: Allergic reactions to measles, mumps, and rubella (MMR) vaccine are rare; some have been attributed to allergy to trace quantities of egg proteins. We report a 17-year-old female who had an anaphylactic reaction to MMR vaccine. A primary vaccination with MMR at age 15 months had been uneventful. She is not allergic to eggs; however, ear and throat pruritus and tongue swelling develop after she eats gelatin. MMR vaccine contains gelatin as a stabilizer. Methods and Results: Prick skin tests were positive to 1 : 10 wt/vol dilutions of MMR vaccine and gelatin but negative to egg. By immunoassay, her serum IgE antibodies were elevated to both MMR vaccine and gelatin, but not to isolated MMR antigens. IgE binding to the gelatin carrier could be inhibited in a dose-dependent fashion by addition of not only MMR vaccine but also gelatin from a variety of animal sources. Immunoblotting confirmed the presence of IgE antibodies to multiple gelatin components as well as to MMR vaccine components. Conclusions: We conclude that the patient has an anaphylactic sensitivity to gelatin, and that her anaphylaxis to MMR vaccine was caused by the gelatin component. This sensitivity may explain other cases of MMR anaphylaxis.

Effect of Parathyroid Hormone on Total Phosphate and Inorganic Phosphate in Blood, Plasma, and Urine of Domestic Fowl

Previous work has shown that parathyroid hormone causes the kidneys of domestic fowl to secrete inorganic phosphate (Pi) into the urine. Secreted Pi is not derived directly from plasma Pi. The objective of the present study was to determine if direct or indirect correlations exist between the urinary Pi excretion rate and the concentration of phosphate in plasma or whole blood. Parathyroid hormone dissolved in gelatin carrier vehicle was injected into immature domestic fowl to cause net renal Pi secretion. Control birds were injected with carrier vehicle alone, resulting in net Pi reabsorption. Urine, plasma, and whole blood samples obtained during net Pi secretion were compared with samples obtained during net Pi reabsorption. Each sample was assayed for total and inorganic phosphate. Neither time course nor point by point comparisons of individual samples provided any evidence that secreted Pi is derived from organic phosphate in plasma or whole blood. These experiments suggest that phosphate pools within the kidneys may serve as a source for the Pi that is secreted in response to parathyroid hormone.

The effect of exogenous acth and pen-confinement on estrous cycle length, plasma steroid levels and ovarian function of beef heifers

Each of 24 pasture-reared crossbred beef heifers were herded from a large pasture on day 14 of the estrous cycle and assigned randomly to one of four treatment groups as follows: Field Control (FC), Field ACTH (FA), Pen Control (PC) and Pen ACTH (PA). Field groups were maintained in a small field, and pen groups were confined in a pen in a pole barn. ACTH groups received 200 IU of ACTH IM daily for days 17 through 21 of the cycle and control groups received only the gelatin carrier IM on the same cycle days. Average cycle lengths for FA and PA heifers were 25 and 24.3 days with an average period from plasma progesterone decline below 1 ng/ml to estrus of 5.5 days. During the ACTH injection period, follicular growth was suppressed and the proestrus plasma estrogen rise was delayed. Average cycle lengths for FC and PC heifers were 20.8 and 22.8 days respectively. All control group heifers exhibited estrus within 2 days of the plasma progesterone decline below 1 ng/ml. In addition, pen confinement heifers showed a trend for extended luteal function and consequent extended estrous cycle length.

Comparison of once daily and twice daily FSH injections for superovulating beef cattle

Twelve cycling Angus-based crossbred cows were used in a crossover experimental design to evaluate two different injection schedules using Follicle Stimulating Hormone (FSH) for superovulating donor cattle. Females randomly assigned to Treatment (A) were given twice daily FSH injections of 5 mg each (12 hours apart) for five consecutive days starting on day 10 of the estrous cycle while those in Treatment (B) received the same daily dose level of FSH, except it was given in a 3.2% protein gelatin carrier vehicle and administered on a once daily injection schedule. Animals in both Treatments (A) and (B) were each given a 30 mg dose of commercially available prostaglandin-F 2α agent 48 hours after the first FSH injection. Cows in estrus were initially handmated to a fertile bull then artificially inseminated 12 hours later with two units of frozen semen. All 12 animals (100%) given twice daily FSH injections and 11 of the females (91.6%) administered once daily FSH injections exhibited standing estrus within 5 days following injection of the luteolytic agent. On day 7 or 8 after the onset of standing estrus a laparotomy was performed to observe ovarian structures. When the superovulation response was evaluated, the mean number of corpora lutea per ovary ranged from 2.9 in the twice daily injection group to 4.1 in the once daily injected group. Unexpectedly, the once daily treated group had significantly more corpora lutea per animal (8.1 vs. 6.4) than those in the twice daily treated group. In addition, mean ovarian size score per animal increased significantly when pre-treatment scores were compared to those recorded following FSH treatment (laparotomy) in both Treatment (A) and (B), however, the post-treatment ovarian size scores were not different between these groups. When evaluating post-treatment follicular development, the once daily injection group had significantly more smaller follicles (<10 mm) and a greater number of ovulatory size follicles (>10 mm) than the twice daily injection group. Furthermore, viable appearing embryos were recovered from both treatment groups and no adverse reactions were observed with the gelatin carrier vehicle in Treatment (B). Since the once daily FSH injection schedule resulted in a superovulatory response equal to or greater than the twice daily FSH injection schedule, this approach to superovulation should not be overlooked by those involved in bovine embryo transplantation.

The effect of porcine calcitonin on plasma calcium in Channa punctatm Bloch

The effect of porcine calcitonin on the plasma calcium of the freshwater Indian murrel, Channa punctatus Bloch has been studied. An injection of 200 MRC mU of porcine calcitonin was given intraperitoneally, while control fish received the gelatin carrier. A significant hypocalcaemia was noted I h after calcitonin treatment (P &lt; 0.001). The plasma calcium had returned to the normal level by the 6th h.