Enzymatic Degradation Analysis Research Articles

On the Effectiveness of Oxygen Plasma and Alkali Surface Treatments to Modify the Properties of Polylactic Acid Scaffolds.

Surface modification of 3D-printed PLA structures is a major issue in terms of increasing the biofunctionality and expanding the tissue engineering applications of these parts. In this paper, different exposure times were used for low-pressure oxygen plasma applied to PLA 3D-printed scaffolds. Alkali surface treatments were also evaluated, aiming to compare the modifications introduced on the surface properties by each strategy. Surface-treated samples were characterized through the quantification of carboxyl groups, energy-dispersive X-ray spectroscopy, water contact angle measurements, and differential scanning calorimetry analysis. The change in the surface properties was studied over a two-week period. In addition, an enzymatic degradation analysis was carried out to evaluate the effect of the surface treatments on the degradation profile of the 3D structures. The physicochemical characterization results suggest different mechanism pathways for each type of treatment. Alkali-treated scaffolds showed a higher concentration of carboxyl groups on their surface, which enhanced the enzymatic degradation rate, but were also proven to be more aggressive towards 3D-printed structures. In contrast, the application of the plasma treatments led to an increased hydrophilicity of the PLA surface without affecting the bulk properties. However, the changes on the properties were less steady over time.

Investigating the Biodegradability and Physical Properties of Starch Derived Bioplastic Films Reinforced with Nanosilica

This study is a pilot investigation on the effect of using nanosilica for reinforcing thermoplastic starch-based bioplastic films. An arbitrary 0.2[Formula: see text]wt.% of nanosilica particles were used to reinforce starch derived bioplastic materials and were further investigated for potential benefits. Nanosilica was extracted from rice husk and was characterized using methods like Fourier transform infrared spectroscopy (FTIR) technique and Brunauer–Emmett–Teller (BET) method. Transmission electron microscopy (TEM) and X-ray diffraction (XRD) techniques were used to determine the structure of nanosilica crystals. Scanning electron microscopy (SEM) technique was used to study the surface topography and composition of nano ‘silica. Both raw and reinforced bioplastic films were tested for thermal stability using thermo gravimetric analysis (TGA) and differential scanning calorimetry (DSC) tests and their performance was compared. Mechanical properties were compared using tensile and tear tests and biodegradability was assessed through enzymatic degradation analysis. It was found that the presence of nanosilica improved the bonding of polymer matrix and in turn increased the thermal stability and tear strength. Nanosilica reinforced matrix resulted in the increase of surface area than raw bioplastic matrix, which lead to high rate of enzymatic reactivity and degradation rate.

Identification and characterization of a novel linkage isomerization in the reaction of trans-diamminedichloroplatinum(II) with 5'-d(TCTACGCGTTCT)

The oligonucleotide 5'-d(TCTACGCGTTCT) reacts with trans-diamminedichloroplatinum(II) to yield primarily trans-[Pt(NH3)2[d(TCTACGCGTTCT)-N7-G(6),N7-G(8)]], containing the desired trans-[Pt(NH3)2[d(GCG)]] 1,3-cross-link. A key element of the platination reaction is the use of low pH to suppress coordination at A(4). The product was fully characterized by pH-dependent NMR titrations, enzymatic degradation analysis, and 195Pt NMR spectroscopy. Interestingly, the 1,3-cross-linked adduct is unstable at neutral pH, rearranging unexpectedly to form the linkage isomer trans-[Pt(NH3)2[d-(TCTACGCGTTCT)-N3-C(5),N7-G(8)]]. This rearrangement product is more stable than the initially formed isomer and could be characterized by pH-dependent NMR titrations, enzymatic degradation analysis, liquid secondary ion mass spectrometric analysis of an enzymatically digested fragment, 195Pt NMR spectroscopy, and modified Maxam-Gilbert footprinting experiments. By contrast, the 1,3-intrastrand cross-linked isomer rearranges during the course of both pH titration and enzymatic degradation experiments to form the 1,4-adduct. The equilibrium constant for this rearrangement is approximately 3, favoring the 1,4-adduct. Kinetic studies of the linkage isomerization reaction reveal t1/2 values for the first-order disappearance of the 1,3-intrastrand cross-linked isomer ranging from 129 (at 30 degrees C) to 3.6 h (at 62 degrees C), with activation parameters delta H not equal to = 91 +/- 2 kJ/mol and delta S not equal to = -58 +/- 8 J/(mol.K). Mechanistic implications of these kinetic results as well as the general relevance of this linkage isomerization reaction to platinum-DNA chemistry are briefly discussed.

Synthesis and characterization of trans-diamminedichloroplatinum(II) adducts of d(CCTCGAGTCTCC).cntdot.d(GGAGACTCGAGG)

The reaction of trans-diamminedichloroplatinum(II) (trans-DDP), the inactive isomer of the anticancer drug cisplatin, with the single-stranded deoxydodecanucleotide d(CCTCGAGTCTCC) in aqueous solution at 37 degrees C was monitored by reversed-phase HPLC. Consumption of the dodecamer follows pseudo-first-order reaction kinetics with a rate constant of 1.25 (4) x 10(-4) s-1. Two intermediates, shown to be monofunctional adducts in which Pt is coordinated to the guanine N7 positions, were trapped with NH4(HCO3) and identified by enzymatic degradation analysis. These monofunctional adducts and a third, less abundant, one are rapidly removed from the DNA by thiourea under mild conditions. When allowed to react further, the monofunctional intermediates formed a single main product that was characterized by 1H NMR spectroscopy and enzymatic digestion as the bifunctional 1,3-intrastrand cross-link trans-[Pt(NH3)2[d(CCTCGAGTCTCC)-N7-G(5),N7-G(7]]). Binding of the trans-[Pt(NH3)2]2+ moiety to the guanosine N7 positions decreases the pKa at N1 and leads to destacking of the intervening A(6) base. The double-stranded trans-DDP-modified and unmodified DNAs were obtained by annealing the complementary strand to the corresponding single strands and then studied by 31P and 1H NMR and UV spectroscopy. trans-DDP binding does not induce large changes in the O-P-O bond or torsional angles of the phosphodiester linkages in the duplex, nor does it significantly alter the UV melting temperature. trans-DDP binding does, however, cause the imino protons of the platinated duplex to exchange rapidly with solvent by 50 degrees C, a phenomenon that occurs at 65 degrees C for the unmodified duplex. A structural model for the platinated double-stranded oligonucleotide was generated through molecular dynamics calculations. This model reveals that the trans-DDP bifunctional adduct can be accommodated within the double helix with minimal distortion of the O-P-O angles and only local disruption of base pairing and destacking of the platinated bases. The model also predicts hydrogen bond formation involving coordinated ammine ligands that bridge the two strands.

Preparation and characterization of new analogues of Dalargin

Tert-butyloxycarbonyl-O-tert-butyloxycarbonyl-L-tyrosyl-D-alanyl-glycyl-L-phenylalanyl-DL-neopentylglycyl-L-arginine was prepared by means of fragment condensation in solution. Protecting groups were removed with trifluoroacetic acid and the diastereoisomeric peptides were separated employing high-performance liquid chromatography (HPLC) and the absolute configuration of neopentylglycine in each diastereomer was determined by two different methods: analysis of enzymatic degradation with carboxypeptidase B and 1H NMR spectroscopic data. The hexapeptide with L-neopentylglycine in position 5 exhibited a 20 times higher activity in a standard test on isolated guinea-pig ileum (GPI). The hexapeptide with D-neopentylglycine in position 5 exhibited a 4 times higher activity than Dalargin in the GPI test but its effect was prolonged in comparison to the analog with L-neopentylglycine and Dalargin.