PubMed · 13273693
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O CHEVITARESE, R F DE PAIVA. [Alginates].. https://pubmed.ncbi.nlm.nih.gov/13273693/
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Keeping equal the initial power drawn (0.27 W l(-1)) in shake flasks and in a stirred fermentor did not reproduce the behaviour of alginate production by Azotobacter vinelandii. A lower mean molecular weight (1.1x10(6) Da) of the polymer was obtained in the bioreactor as compared to that obtained in shake flasks (1.9x10(6) Da). The reasons for this can reside in the fact that the evolution of the power drawn in the shake flasks could be considerably different to that observed in the stirred bioreactor. A drastic drop in the specific power drawn is expected in the shake flasks as a consequence of the increased viscosity, which caused the liquid not following the movement of the shaker. This was supported by the fact that cultures developed in the fermentor at lower initial power drawn (as low as 0.027-0.056 W l(-1)) or in a culture in which the power drawn was deliberately reduced along cultivation, produced alginates with similar molecular characteristics as that obtained in shake flasks.
Bone morphogenetic protein-2 (BMP-2) promotes the formation and regeneration of bone and cartilage, and also participates in organogenesis, cell differentiation, cell proliferation, and apoptosis. BMP-2 has two epitopes referred to as the "wrist epitope" and the "knuckle epitope". The wrist epitope is thought to bind to BMP receptor IA and the knuckle epitope to BMP receptor type II. However, the precise receptor-binding region in BMP-2 has not yet been clarified. Here, we report that a synthetic peptide, KIPKASSVPTELSAISTLYL, corresponding to residues 73-92 of the knuckle epitope of BMP-2, elevated alkaline phosphatase (ALP) activity in the murine multipotent mesenchymal cell line, C3H10T1/2. The 73-92 peptide significantly inhibited the binding of rhBMP-2 to both BMP receptors type IA and type II. The 73-92 peptide also promoted the expression of osteocalcin mRNA and induced ectopic calcification when it was immobilized on a covalently cross-linked alginate gel and implanted into rat calf muscle. The X-ray diffraction (XRD) pattern of the calcified product was identical to that of the rat tibia, and the major peaks were attributed to hydroxyapatite. These results indicate that the 73-92 peptide may be one of the receptor-binding sites on BMP-2 and may stimulate bone precursor cells to induce calcification.
The entrapment of enzymes, drugs, cells or tissue fragments in alginates cross-linked with Ca(2+) or Ba(2+) has great potential in basic research, biotechnology and medicine. The swelling properties and, in turn, the mechanical stability are key factors in designing an optimally cross-linked hydrogel matrix. These parameters depend critically on the cross-linking process and seemingly minor modifications in manufacture have a large impact. Thus, sensitive and non-invasive tools are required to determine the spatial homogeneity and efficacy of the cross-linking process. Here, we show for alginate microcapsules (between 400 microm and 600 microm in diameter) that advanced (1)H NMR imaging, along with paramagnetic Cu(2+) as contrast agent, can be used to validate the cross-linking process. Two- and three-dimensional images and maps of the spin-lattice relaxation time T(1) of Ba(2+) cross-linked microcapsules exposed to external Cu(2+) yielded qualitative as well as quantitative information about the accumulation of Cu(2+) within and removal from microcapsules upon washing with Cu(2+) free saline solution. The use of Cu(2+) (having a slightly higher affinity constant to alginate than Ba(2+)) for gelling gave a complementary insight into the spatial homogeneity of the cross-linking process together with information about the mechanical stability of the microcapsules. The potential of this technique was demonstrated for alginates extracted from two different algal sources and cross-linked either externally by the conventional air-jet dropping method or internally by the "crystal gun" method.