Preparation and in vitro release profiling of PLGA microspheres containing BSA as a model protein

Conventional drug formulations are incapable of adequate delivery of proteins and peptides for therapeutic purposes. As these molecules have very short biological half-life, multiple dosing is required to achieve the desirable therapeutic effects. Microspheres are able to encapsulate proteins and peptide in the polymeric matrix while protecting them from enzymatic degradation. In this study Bovine Serum Albumin (BSA) matrix type microspheres were fabricated using Polylactide-co-glycolide (PLGA) by double emulsion solvent evaporation method. The effects of variables such as homogenizer speed, molecular weight of polymer and the effect of pH of the water phases, were investigated against factors such as drug loading, encapsulation efficiency, morphology, size, drug distribution and release profile of the microspheres. Results, suggested that an increase in homogenization speed leads to a decrease in microsphere size. The increase in homogenization speed also caused a significant effect on the release profile only when higher molecular weight of polymer had been used.. The pH change of the internal aqueous phase led to modification of surface morphology of spheres to a porous structure that significantly increased the total amount of released protein. Integrity of protein structure was intact as shown by SDS-PAGE. According to the results, it can be concluded that we achieved a reproducible method regarding controlled protein delivery for different sizes of particles.

Absorção de IgG via colostro em leitões biológicos e adotados após a uniformização da leitegada / Absorption of IgG via colostrum in biological piglets and adopted piglets after crossfostering

Nove fêmeas de quinto parto (OP5) foram imunizadas com 4mg e 2mg de albumina sérica bovina (BSA) aos 70 e 100 dias de gestação, respectivamente. A uniformização da leitegada foi realizada 4,9±1,9h após o nascimento, antes de os leitões efetuarem a primeira mamada. As leitegadas foram compostas por cinco leitões biológicos (LB) e cinco leitões adotados (LA), com pesos semelhantes ao nascimento. Foram coletadas amostras de sangue dos leitões ao nascimento e 24h após, das fêmeas ao pós-parto e de colostro de cada grupo de tetos ao parto e 24h após. As amostras de soro e colostro foram quantificadas para IgG pelo ELISA indireto. A densidade ótica de IgG anti-BSA (DOIgG-BSA) dos leitões (24h de vida) foi correlacionada com a das fêmeas. A DOIgG-BSA entre LB e LA foi semelhante, assim como entre os grupos de tetos, ao parto e 24h após. Entretanto, ocorreu redução na DOIgG-BSA do parto até 24h após. LB e LA absorveram a mesma quantidade de IgG via colostro, quando a uniformização foi realizada até 5h pós-parto, independentemente do teto em que os leitões mamaram, uma vez que esses possuem a mesma concentração de IgG.

Nine sows of fifth parity (PO5) were immunized with 4mg and 2mg of bovine serum albumin (BSA) at 70 and 100d of gestation, respectively. Cross fostering was performed 4.9±1.9h after birth, before piglets had their first suckling. Litters were composed of five biological piglets (BP) and five adopted piglets (AP), with similar weight at birth. Blood samples were collected from piglets (at birth and at 24h of life) and from females (after farrowing) and colostrum from each group of teats (at farrowing time and after 24h). Samples of serum and colostrum were quantified to IgG by indirect ELISA. Optical density of IgG anti-BSA (ODIgG-BSA) from piglets (24h of life) was correlated with dams. ODIgG-BSA was similar among BP and AP, as well as among pairs of teats (at farrowing time and after 24h). However, there was a decrease in ODIgG-BSA from farrowing up to 24h after birth. BP and AP absorbed the same amount of IgG via colostrum, when cross fostering was evaluated 5h after farrowing, regardless of the teat suckled, since these have the same concentration of IgG.

Effects of soluble antigen-induced immune cell activation on steroidogenesis in murine lymphoid organ.

The effect of soluble antigenic (bovine serum albumin, BSA) stimulation to induce steroidogenesis in murine lymphoid organs with concomitant changes in proinflammatory or inflammatory cytokine levels and its implication in the alteration of T-cell response was studied in the mice. Male Swiss albino mice (6-8 weeks old) with average body weight (20 +/- 4 g) were randomly assigned to 3 groups and injected with BSA in presence and absence of Freund's complete or incomplete adjuvant, whereas the control group received only saline. After 3 weeks, animals were sacrificed, and serums as well as lymphoid organs were collected. From the lymphoid tissue homogenate, the activities of steroidogenic enzymes and corticosterone and cytokine levels of the serum were estimated. Steroidogenic enzyme activities in murine lymphoid organs, as well as the pro-inflammatory and inflammatory cytokines levels in serum increased after Freund's complete adjuvant-emulsified BSA administration, as compared to control. The serum corticosterone and serum cytokine profile were also elevated. Results suggested that soluble protein antigen (BSA) administration stimulated steroidogenesis in murine lymphoid tissues and rise in the pro-inflammatory or inflammatory cytokine levels might indicate monocyte recruitment as well as TH1 activation.

An Experimental Study on Immune Complex Induced Athritis in Rabbits: Reference to Macrophages and M-type Cells of the Synovium

This study evaluates the pathogenesis of rheumatoid arthritis by producing immune complex induced arthritis with an intra-articular injection of BSA in immunized rabbits, and the effect of systemic administration of cyclophosphamide and local administration of anti-macrophage serum. The reduction of inflammatory reaction by cyclophosphamide administration appears to be caused mainly by selective depletion of the neutrophils, and partly by immune suppression. It appears that the rabbit abdominal macrophage has the common morphologic, functional and antigenic patterns with the M-type synovial lining cells. There is another possibility that the cross-reacting antigens between macrophage and the M-type cell of the synovial lining may exist. It is concluded that in this experimental immune complex arthritis, the site of localization of immune complexes seems to be the synovial, M-type cell, and the tissue injury of synovium is largely mediated not only by neutrophils and complement, but also by macrophages.