Nano-yarn carbon nanotube fiber based enzymatic glucose biosensor

This is the author's accepted manuscript. The final published article is available from the link below. Copyright @ 2010 IOP Publishing Ltd.A novel brush-like electrode based on carbon nanotube (CNT) nano-yarn fiber has been designed for electrochemical biosensor applications and its efficacy as an enzymatic glucose biosensor demonstrated. The CNT nano-yarn fiber was spun directly from a chemical-vapor-deposition (CVD) gas flow reaction using a mixture of ethanol and acetone as the carbon source and an iron nano-catalyst. The fiber, 28 µm in diameter, was made of bundles of double walled CNTs (DWNTs) concentrically compacted into multiple layers forming a nano-porous network structure. Cyclic voltammetry study revealed a superior electrocatalytic activity for CNT fiber compared to the traditional Pt–Ir coil electrode. The electrode end tip of the CNT fiber was freeze-fractured to obtain a unique brush-like nano-structure resembling a scale-down electrical 'flex', where glucose oxidase (GOx) enzyme was immobilized using glutaraldehyde crosslinking in the presence of bovine serum albumin (BSA). An outer epoxy-polyurethane (EPU) layer was used as semi-permeable membrane. The sensor function was tested against a standard reference electrode. The sensitivities, linear detection range and linearity for detecting glucose for the miniature CNT fiber electrode were better than that reported for a Pt–Ir coil electrode. Thermal annealing of the CNT fiber at 250 °C for 30 min prior to fabrication of the sensor resulted in a 7.5 fold increase in glucose sensitivity. The as-spun CNT fiber based glucose biosensor was shown to be stable for up to 70 days. In addition, gold coating of the electrode connecting end of the CNT fiber resulted in extending the glucose detection limit to 25 µM. To conclude, superior efficiency of CNT fiber for glucose biosensing was demonstrated compared to a traditional Pt–Ir sensor.Brunel University, the Royal Society and the National Institute of Health

Cross-linked cholecyst-derived extracellular matrix for abdominal wall repair

Abdominal wall repair frequently utilizes either non-degradable or bio-degradable meshes, which are found to stimulate undesirable biological tissue responses or which possess suboptimal degradation rate. In this study, a biologic mesh prototype made from carbodiimide-cross-linked cholecyst-derived extracellular matrix (EDCxCEM) was compared with small intestinal submucosa (Surgisis®), cross-linked bovine pericardium (Peri-Guard®), and polypropylene (Prolene®) meshes in an in vivo rabbit model. The macroscopic appearance and stereological parameters of the meshes were evaluated. Tailoring the degradation of the EDCxCEM mesh prevents untimely degradation, while allowing cellular infiltration and mesh remodelling to take place in a slower but predictable manner. The results suggest that the cross-linked biodegradable cholecyst-derived biologic mesh results in no seroma formation, low adhesion, and moderate stretching of the mesh. In contrast to Surgisis®, Peri-Guard®, and Prolene® meshes, the EDCxCEM mesh showed a statistically significant increase in the volume fraction (Vv) of collagen (from 34% to 52.1%) in the central fibrous tissue region at both day 28 and day 56. The statistically high Length density (Lv), of blood vessels for the EDCxCEM mesh at 28 days was reflected also by the higher cellular activity (high Vv of fibroblast and moderate Vv of nuclei) indicating remodelling of this region in the vicinity of a slowly degrading EDCxCEM mesh. The lack of mesh area stretching/ shrinkage in the EDCxCEM mesh showed that the remodelled tissue was adequate to prevent hernia formation. The stereo-histological assays suggest that the EDCxCEM delayed degradation profile supports host wound healing processes including collagen formation, cellular infiltration, and angiogenesis. The use of cross-linked cholecyst-derived extracellular matrix for abdominal wall repair is promising

Buttressing staples with cholecyst-derived extracellular matrix (CEM) reinforces staple lines in an ex vivo peristaltic inflation model

This is the author's accepted manuscript. The final published article is available from the link below. Copyright @ Springer Science + Business Media, LLC 2008Background - Staple line leakage and bleeding are the most common problems associated with the use of surgical staplers for gastrointestinal resection and anastomotic procedures. These complications can be reduced by reinforcing the staple lines with buttressing materials. The current study reports the potential use of cholecyst-derived extracellular matrix (CEM) in non-crosslinked (NCEM) and crosslinked (XCEM) forms, and compares their mechanical performance with clinically available buttress materials [small intestinal submucosa (SIS) and bovine pericardium (BP)] in an ex vivo small intestine model. Methods - Three crosslinked CEM variants (XCEM0005, XCEM001, and XCEM0033) with different degree of crosslinking were produced. An ex vivo peristaltic inflation model was established. Porcine small intestine segments were stapled on one end, using buttressed or non-buttressed surgical staplers. The opened, non-stapled ends were connected to a peristaltic pump and pressure transducer and sealed. The staple lines were then exposed to increased intraluminal pressure in a peristaltic manner. Both the leak and burst pressures of the test specimens were recorded. Results - The leak pressures observed for non-crosslinked NCEM (137.8 ± 22.3 mmHg), crosslinked XCEM0005 (109.1 ± 14.1 mmHg), XCEM001 (150.1 ± 16.0 mmHg), XCEM0033 (98.8 ± 10.5 mmHg) reinforced staple lines were significantly higher when compared to non-buttressed control (28.3 ± 10.8 mmHg) and SIS (one and four layers) (62.6 ± 11.8 and 57.6 ± 12.3 mmHg, respectively) buttressed staple lines. NCEM and XCEM were comparable to that observed for BP buttressed staple lines (138.8 ± 3.6 mmHg). Only specimens with reinforced staple lines were able to achieve high intraluminal pressures (ruptured at the intestinal mesentery), indicating that buttress reinforcements were able to withstand pressure higher than that of natural tissue (physiological failure). Conclusions - These findings suggest that the use of CEM and XCEM as buttressing materials is associated with reinforced staple lines and increased leak pressures when compared to non-buttressed staple lines. CEM and XCEM were found to perform comparably with clinically available buttress materials in this ex vivo model.Enterprise Irelan

Nanomaterials in glucose sensing

Research shows that overall, nanomaterials help address the problems with conventional optical and electrochemical biosensors, by enhancing the preferential detection of glucose or its oxidation products through better electron transfer kinetics, sensitivity and response time, while lowering the operating over-voltages for energy efficiency and avoid interference. The reproducible production of nano-materials and nano-structures at low cost is vital for the successful development of nano-technologies for glucose sensing. Several products, especially, home glucose monitoring devices, use nano-materials, but the need for reliable long-term CGM is still unmet. Nano-materials and nano-technologies have an important role in achieving the long-awaited CGM technology

Buttressing staples with cholecyst-derived extracellular matrix (cem) reinforces staple lines in an ex vivo peristaltic inflation model

Background Staple line leakage and bleeding are the most common problems associated with the use of surgical staplers for gastrointestinal resection and anastomotic procedures. These complications can be reduced by reinforcing the staple lines with buttressing materials. The current study reports the potential use of cholecyst-derived extracellular matrix (CEM) in non-crosslinked (NCEM) and crosslinked (XCEM) forms, and compares their mechanical performance with clinically available buttress materials [small intestinal submucosa (SIS) and bovine pericardium (BP)] in an ex vivo small intestine model. Methods Three crosslinked CEM variants (XCEM0005, XCEM001, and XCEM0033) with different degree of crosslinking were produced. An ex vivo peristaltic inflation model was established. Porcine small intestine segments were stapled on one end, using buttressed or non-buttressed surgical staplers. The opened, non-stapled ends were connected to a peristaltic pump and pressure transducer and sealed. The staple lines were then exposed to increased intraluminal pressure in a peristaltic manner. Both the leak and burst pressures of the test specimens were recorded. Results The leak pressures observed for non-crosslinked NCEM (137.8 +/- 22.3 mmHg), crosslinked XCEM0005 (109.1 +/- 14.1 mmHg), XCEM001 (150.1 +/- 16.0 mmHg), XCEM0033 (98.8 +/- 10.5 mmHg) reinforced staple lines were significantly higher when compared to non-buttressed control (28.3 +/- 10.8 mmHg) and SIS (one and four layers) (62.6 +/- 11.8 and 57.6 +/- 12.3 mmHg, respectively) buttressed staple lines. NCEM and XCEM were comparable to that observed for BP buttressed staple lines (138.8 +/- 3.6 mmHg). Only specimens with reinforced staple lines were able to achieve high intraluminal pressures (ruptured at the intestinal mesentery), indicating that buttress reinforcements were able to withstand pressure higher than that of natural tissue (physiological failure). Conclusions These findings suggest that the use of CEM and XCEM as buttressing materials is associated with reinforced staple lines and increased leak pressures when compared to non-buttressed staple lines. CEM and XCEM were found to perform comparably with clinically available buttress materials in this ex vivo model

Efficacy of crosslinking on tailoring in vivo biodegradability of fibro-porous decellularized extracellular matrix and restoration of native tissue structure: a quantitative study using stereology methods

Cholecyst-derived extracellular matrix (CEM) is a fibro-porous decellularized serosal layer of porcine gall-bladder. CEM loses 90% of its weight at 48h of in vitro collagenase digestion, but takes two months to be completely resorbed in vivo. Carbodiimide (EDC) crosslinking helps tailoring CEM\u27s in vitro collagenase susceptibility. Here, the efficacy of EDC crosslinking on tailoring in vivo biodegradability of CEM is reported. CEM crosslinked with 0.0005 and 0.0033x10(3) M of EDC/mg that lose 80% and 0% of their weight respectively to in vitro collagenase digestion, were present even after 180 days in vivo. Quantitative histopathology using stereology methods confirmed our qualitative observation that even a tiny degree of crosslinking can significantly prolong the rate of in vivo degradation and removal of CEM

A clinically relevant in vivo model for the assessment of scaffold efficacy in abdominal wall reconstruction

An animal model that allows for assessment of the degree of stretching or contraction of the implant area and the in vivo degradation properties of biological meshes is required to evaluate their performance in vivo. Adult New Zealand rabbits underwent full thickness subtotal unilateral rectus abdominis muscle excision and were reconstructed with the nonbiodegradable Peri-Guard (R), Prolene (R) or biodegradable Surgisis (R) meshes. Following 8 weeks of recovery, the anterior abdominal wall tissue samples were collected for measurement of the implant dimensions. The Peri-Guard and Prolene meshes showed a slight and obvious shrinkage, respectively, whereas the Surgisis mesh showed stretching, resulting in hernia formation. Surgisis meshes showed in vivo biodegradation and increased collagen formation. This surgical rabbit model for abdominal wall defects is advantageous for evaluating the in vivo behaviour of surgical meshes. Implant area stretching and shrinkage were detected corresponding to mesh properties, and histological analysis and stereological methods supported these findings

Amine functionalization of cholecyst-derived extracellular matrix with generation 1 pamam dendrimer

A method to functionalize cholecyst-derived extracellular matrix (CEM) with free amine groups was established in an attempt to improve its potential for tethering of bioactive molecules. CEM was incorporated with Generation-1 polyamidoamine (G1 PAMAM) dendrimer by using N-(3-dimethylaminopropyl)-N\u27-ethylcarbodiimide and N-hydroxysuccinimide cross-linking system. The nature of incorporation of PAMAM dendrimer was evaluated using shrink temperature measurements, Fourier transform infrared (FTIR) assessment, ninhydrin assay, and swellability. The effects of PAMAM incorporation on mechanical and degradation properties of CEM were evaluated using a uniaxial mechanical test and collagenase degradation assay, respectively. Ninhydrin assay and FTIR assessment confirmed the presence of increasing free amine groups with increasing quantity of PAMAM in dendrimer-incorporated CEM (DENCEM) scaffolds. The amount of dendrimer used was found to be critical in controlling scaffold degradation, shrink temperature, and free amine content. Cell culture studies showed that fibroblasts seeded on DENCEM maintained their metabolic activity and ability to proliferate in vitro. In addition, fluorescence cell staining and scanning electron microscopy analysis of cell-seeded DENCEM showed preservation of normal fibroblast morphology and phenotype