Understanding the relationships between material surface properties, adsorbed proteins, and cellular responses is essential to designing optimal material surfaces for implantation and tissue engineering. In this study, we have prepared model surfaces with different functional groups to provide a range of surface wettability and charge. The cellular responses of attachment, spreading, and cytoskeletal organization have been studied following preadsorption of these surfaces with dilute serum, specific serum proteins, and individual components of the extracellular matrix. When preadsorbed with dilute serum, cell attachment, spreading, and cytoskeletal organization were significantly greater on hydrophilic surfaces relative to hydrophobic surfaces. Among the hydrophilic surfaces, differences in charge and wettability influenced cell attachment but not cell area, shape, or cytoskeletal organization. Moderately hydrophilic surfaces (20-40 degree water contact angle) promoted the highest levels of cell attachment. Preadsorption of the model surfaces with bovine serum albumin (BSA) resulted in a pattern of cell attachment very similar to that observed following preadsorption with dilute serum, suggesting an important role for BSA in regulating cell attachment to biomaterials exposed to complex biological media.
Over the last 50 yr, the study of intestinal peptide transport has rapidly evolved into a field with exciting nutritional and biomedical applications. In this review, we describe from a historical and current perspective intestinal peptide transport, the importance of peptides to whole-body nutrition, and the cloning and characterization of the intestinal peptide transporter, PepT1. We focus on the nutritional significance of peptide transport and relate these findings to livestock and poultry. Amino acids are transported into the enterocyte as free AA by a variety of AA transporters that vary in substrate specificity or as di- and tripeptides by the peptide transporter, PepT1. Expression of PepT1 is largely restricted to the small intestine in most species; however, in ruminants, peptide transport and activity is observed in the rumen and omasum. The extent to which peptides are absorbed and utilized is still unclear. In ruminants, peptides make a contribution to the portal-drained visceral flux of total AA and are detected in circulating plasma. Peptides can be utilized by the mammary gland for milk protein synthesis and by a variety of other tissues. We discuss the factors known to regulate expression of PepT1 including development, diet, hormones, diurnal rhythm, and disease. Expression of PepT1 is detected during embryological stages in both birds and mammals and increases with age, a strategic event that allows for the immediate uptake of nutrients after hatch or birth. Both increasing levels of protein in the diet and dietary protein deficiencies are found to upregulate the peptide transporter. We also include in this review a discussion of the use of dietary peptides and potential alternate routes of nutrient delivery to the cell. Our goal is to impart to the reader the nutritional implications of peptide transport and dietary peptides and share discoveries that shed light on various biological processes, including rapid establishment of intestinal function in early neonates and maintenance of intestinal function during fasting, starvation, and disease states.
The objective of this study was to investigate intestinal nutrient transporter and enzyme mRNA in broilers selected on corn- and soybean-based (line A) or wheat-based (line B) diets. We investigated the peptide transporter PepT1, 10 amino acid transporters (rBAT, b(o,+)AT, ATB(o,+), CAT1, CAT2, LAT1, y(+)LAT1, y(+)LAT2, B(o)AT, and EAAT3), 4 sugar transporters (SGLT1, SGLT5, GLUT5, and GLUT2), and a digestive enzyme (aminopeptidase N). Intestine was collected at embryo d 18 and 20, day of hatch, and d 1, 3, 7, and 14 posthatch. The mRNA abundance of each gene was assayed using real-time PCR and the absolute quantification method. For PepT1, line B had greater quantities of mRNA compared with line A (P = 0.001), suggesting a greater capacity for absorption of amino acids as peptides. Levels of PepT1 mRNA were greatest in the duodenum (P < 0.05), whereas the abundances of SGLT1, GLUT5, and GLUT2 mRNA were greatest in the jejunum (P < 0.05). Abundances of EAAT3, b(o,+)AT, rBAT, B(o)AT, LAT1, CAT2, SGLT5, and aminopeptidase N mRNA were greatest in the ileum (P < 0.05). Quantities of PepT1, EAAT3, B(o)AT, SGLT1, GLUT5, and GLUT2 mRNA increased linearly (P < 0.01), whereas CAT1, CAT2, y(+)LAT1, and LAT1 mRNA decreased linearly (P < 0.05) with age. Abundance of y(+)LAT2 mRNA changed cubically (P = 0.002) with peaks of expression at day of hatch and d 7, and aminopeptidase N and SGLT5 mRNA changed quadratically (P = 0.005) with age. These results provide a comprehensive profile of the temporal and spatial expression of nutrient transporter mRNA in the small intestine of chicks.
Many research and commercial applications use a synthetic substrate which is seeded with cells in a serum-containing medium. The surface properties of the material influence the composition of the adsorbed protein layer, which subsequently regulates a variety of cell behaviors such as attachment, spreading, proliferation, migration, and differentiation. In this study, we examined the relationships among cell attachment, spreading, cytoskeletal organization, and migration rate for MC3T3-E1 osteoblasts on glass surfaces modified with -SO(x), -NH(2), -N(+)(CH(3))(3), -SH, and -CH(3) terminal silanes. We also studied the relationship between cell spread area and migration rate for a variety of anchorage-dependent cell types on a model polymeric biomaterial, poly(acrylonitrile-vinylchloride). Our results indicated that MC3T3-E1 osteoblast behavior was surface chemistry dependent, and varied with individual functional groups rather than general surface properties such as wettability. In addition, cell migration rate was inversely related to cell spread area for MC3T3-E1 osteoblasts on a variety of silane-modified surfaces as well as for different anchorage-dependent cell types on a model polymeric biomaterial. Furthermore, the data revealed significant differences in migration rate among different cell types on a common polymeric substrate, suggesting that cell type-specific differences must be considered when using, selecting, or designing a substrate for research and therapeutic applications.
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