Biomineralization potential and cellular response of PHB and PHBV blends with natural anionic polysaccharides.

In this paper, the biomineralization potential and cellular response of novel blend films of the anionic sulfated polysaccharides kappa-carrageenan (KCG) and fucoidan (FUC) derived from seaweeds with semi-crystalline polyhydroxybutyrate (PHB) and polyhydroxybutyrate-co-valerate (PHBV), respectively, were analyzed. The incorporation of KCG and FUC into PHB and PHBV, which has been studied here for the first time, led to an overall decrease in crystallinity, enhanced surface hydrophilicity, reduced brittleness and faster degradation of the polymer blend films. All PHB/KCG, PHBV/KCG and PHBV/FUC films exhibited a two-stage mass loss profiles with pH stabilization. PHBV/KCG film showed the highest biomineralization activity due the presence of sulfate groups on the surface of the films. NIH3T3 cells attached and proliferated well on all blend films on account of enhanced surface hydrophilicity and improved flexibility. PHBV/KCG led to a promoted cellular activity compared to PHBV/FUC, presumably due to phase separation and higher amount of biopolymer on the film surface that was a consequence of the immiscibility of the polymers in the blend films.

Third generation poly(hydroxyacid) composite scaffolds for tissue engineering.

Bone tissue engineering based on scaffolds is quite a complex process as a whole gamut of criteria needs to be satisfied to promote cellular attachment, proliferation and differentiation: biocompatibility, right surface properties, adequate mechanical performance, controlled bioresorbability, osteoconductivity, angiogenic cues, and vascularization. Third generation scaffolds are more of composite types to maximize biological-mechanical-chemical properties. In the present review, our focus is on the performance of micro-organism-derived polyhydroxyalkanoates (PHAs)-polyhydroxybutyrate (PHB) and polyhydroxybutyrate-co-valerate (PHBV)-composite scaffolds with ceramics and natural polymers for tissue engineering applications with emphasis on bone tissue. We particularly emphasize on how material properties of the composites affect scaffold performance. PHA-based composites have demonstrated their biocompatibility with a range of tissues and their capacity to induce osteogenesis due to their piezoelectric properties. Electrospun PHB/PHBV fiber mesh in combination with human adipose tissue-derived stem cells (hASCs) were shown to improve vascularization in engineered bone tissues. For nerve and skin tissue engineering applications, natural polymers such as collagen and chitosan remain the gold standard but there is scope for development of scaffolds combining PHAs with other natural polymers which can address some of the limitations such as brittleness, lack of bioactivity and slow degradation rate presented by the latter. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 1667-1684, 2017.

The use of NaCl addition for the improvement of polyhydroxyalkanoate production by Cupriavidus necator.

External stress factors in the form of ionic species or temperature increases have been shown to produce a stress response leading to enhanced PHA production. The effect of five different NaCl concentrations, namely 3.5, 6.5, 9, 12 and 15 g/l NaCl on PHA productivity using Cupriavidus necator has been investigated alongside a control (no added NaCl). A dielectric spectroscopy probe was used to measure PHA accumulation online in conjunction with the chemical offline analysis of PHA. The highest PHA production was obtained with the addition of 9 g/l NaCl, which yielded 30% higher PHA than the control. Increasing the addition of NaCl to 15 g/l was found to inhibit the production of PHA. NaCl addition can therefore be used as a simple, low cost, sustainable, non toxic and non reactive external stress strategy for increasing PHA productivity.

Increasing polyhydroxyalkanoate (PHA) yields from Cupriavidus necator by using filtered digestate liquors.

The production of polyhydroxyalkanoates (PHAs) using digestate liquor as culture media is a novel application to extend the existing uses of digestates. In this study, two micro-filtered digestates (0.22 µm) were evaluated as a source of complex culture media for the production of PHA by Cupriavidus necator as compared to a conventional media. Culture media using a mixture of micro-filtered liquors from food waste and from wheat feed digesters showed a maximum PHA accumulation of 12.29 g/l PHA, with 90% cell dry weight and a yield of 0.48 g PHA/g VFA consumed, the highest reported to date for C. necator studies. From the analysis of the starting and residual media, it was concluded that ammonia, potassium, magnesium, sulfate and phosphate provided in the digestate liquors were vital for the initial growth of C. necator whereas copper, iron and nickel may have played a significant role in PHA accumulation.

Addressing the challenge of optimum polyhydroxyalkanoate harvesting: monitoring real time process kinetics and biopolymer accumulation using dielectric spectroscopy.

In this study, dielectric spectroscopy was utilised to evaluate and define the optimum harvesting time for polyhydroxyalkanoates (PHA) production. It is essential to harvest PHA at the optimum time during fermentation for maximum yield, otherwise cells start degrading. Two carbon sources (acetic and butyric acids) were used in laboratory based experiments and a number of samples were measured ex situ for PHA production. The real-time measured capacitance in addition of identifying the cells growth phase, it correlated very well with ex situ measured PHA produced within the cells. The probe has proven to be a useful tool to assess process kinetics, to monitor real-time cell growth, PHA produced and defining the optimum harvesting time.