Designing of Collagen Based Poly(3-hydroxybutyrate-co-4-hydroxybutyrate) Scaffolds for Tissue Engineering

P(3HB-co-4HB) copolymer was modified using collagen by adapting dual solvent system. The surface properties of samples were characterized by Fourier transforminfrared spectroscopy (FTIR), scanning electron microscopy (SEM), organic elemental analysis (CHN analysis), and water contact angle measurements.The effects of collagen concentration, scaffold thickness, and 4HB molar fraction on the hydrophilicitywere optimized by the Taguchi method.The orthogonal array experiment was conducted to obtain the response for a hydrophilic scaffold. Analysis of variance (ANOVA) was used to determine the significant parameters and determine the optimal level for each parameter. The results also showed that the hydrophilicity of P(3HB-co-4HB)/collagen blend scaffolds increased as the collagen concentration increased up to 15 wt% with a molar fraction of 50mol% at 0.1mm scaffold thickness. The biocompatibility of the P(3HB-co-4HB)/collagen blend surface was evaluated by fibroblast cell (L929) culture.The collagen blend scaffold surfaces showed significant cell adhesion and growth as compared to P(3HB-co-4HB) copolymer scaffolds

Elucidating the Surface Functionality of Biomimetic RGD Peptides Immobilized on Nano-P(3HB-co-4HB) for H9c2 Myoblast Cell Proliferation

Biomaterial scaffolds play crucial role to promote cell proliferation and foster the regeneration of new tissues. The progress in material science has paved the way for the generation of ingenious biomaterials. However, these biomaterials require further optimization to be effectively used in existing clinical treatments. It is crucial to develop biomaterials which mimics structure that can be actively involved in delivering signals to cells for the formation of the regenerated tissue. In this research we nanoengineered a functional scaffold to support the proliferation of myoblast cells. Poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] copolymer is chosen as scaffold material owing to its desirable mechanical and physical properties combined with good biocompatibility, thus eliciting appropriate host tissue responses. In this study P(3HB-co-4HB) copolymer was biosynthesized using Cupriavidus malaysiensis USMAA1020 transformant harboring additional PHA synthase gene, and the viability of a novel P(3HB-co-4HB) electrospun nanofiber scaffold, surface functionalized with RGD peptides, was explored. In order to immobilize RGD peptides molecules onto the P(3HB-co-4HB) nanofibers surface, an aminolysis reaction was performed. The nanoengineered scaffolds were characterized using SEM, organic elemental analysis (CHN analysis), FTIR, surface wettability and their in vitro degradation behavior was evaluated. The cell culture study using H9c2 myoblast cells was conducted to assess the in vitro cellular response of the engineered scaffold. Our results demonstrated that nano-P(3HB-co-4HB)-RGD scaffold possessed an average fiber diameter distribution between 200 and 300 nm, closely biomimicking, from a morphological point of view, the structural ECM components, thus acting as potential ECM analogs. This study indicates that the surface conjugation of biomimetic RGD peptide to the nano-P(3HB-co-4HB) fibers increased the surface wettability (15 ± 2°) and enhanced H9c2 myoblast cells attachment and proliferation. In summary, the study reveals that nano-P(3HB-co-4HB)-RGD scaffold can be considered a promising candidate to be further explored as cardiac construct for building cardiac construct

Designing of Collagen Based Poly(3-hydroxybutyrate- co

P(3HB-co-4HB) copolymer was modified using collagen by adapting dual solvent system. The surface properties of samples were characterized by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), organic elemental analysis (CHN analysis), and water contact angle measurements. The effects of collagen concentration, scaffold thickness, and 4HB molar fraction on the hydrophilicity were optimized by the Taguchi method. The orthogonal array experiment was conducted to obtain the response for a hydrophilic scaffold. Analysis of variance (ANOVA) was used to determine the significant parameters and determine the optimal level for each parameter. The results also showed that the hydrophilicity of P(3HB-co-4HB)/collagen blend scaffolds increased as the collagen concentration increased up to 15 wt% with a molar fraction of 50 mol% at 0.1 mm scaffold thickness. The biocompatibility of the P(3HB-co-4HB)/collagen blend surface was evaluated by fibroblast cell (L929) culture. The collagen blend scaffold surfaces showed significant cell adhesion and growth as compared to P(3HB-co-4HB) copolymer scaffolds

Enhanced production of poly (3-hydroxybutyrate-co-4-hydroxybutyrate) copolymer with manipulated variables and its properties

Cupriavidus sp. USMAA1020, a local isolate was able to biosynthesis poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] copolymer with various 4HB precursors as the sole carbon source. Manipulation of the culture conditions such as cell concentration, phosphate ratio and culture aeration significantly affected the synthesis of P(3HB-co-4HB) copolymer and 4HB composition. P(3HB-co-4HB) copolymer with 4HB compositions ranging from 23 to 75 mol% 4HB with various mechanical and thermal properties were successfully produced by varying the medium aeration. The physical and mechanical properties of P(3HB-co-4HB) copolymers were characterized by NMR spectroscopy, gel-permeation chromatography, tensile test, and differential scanning calorimetry. The number-average molecular weights (M (n)) of copolymers ranged from 260 x 10(3) to 590 x 10(3)Da, and the polydispersities (M (w)/M (n)) were between 1.8 and 3.0. Increases in the 4HB composition lowered the molecular weight of these copolymers. In addition, the increase in 4HB composition affected the randomness of copolymer, melting temperature (T (m)), glass transition temperature (T (g)), tensile strength, and elongation to break. Enzymatic degradation of P(3HB-co-4HB) films with an extracellular depolymerase from Ochrobactrum sp. DP5 showed that the degradation rate increased proportionally with time as the 4HB fraction increased from 17 to 50 mol% but were much lower with higher 4HB fraction. Degradation of P(3HB-co-4HB) films with lipase from Chromobacterium viscosum exhibited highest degradation rate at 75 mol% 4HB. The biocompatibility of P(3HB-co-4HB) copolymers were evaluated and these copolymers have been shown to support the growth and proliferation of fibroblast cells

Extracellular Polyhydroxyalkanoate Depolymerase by Acidovorax sp. DP5

Bacteria capable of degrading polyhydroxyalkanoates (PHA) by secreting extracellular depolymerase enzymes were isolated from water and soil samples collected from various environments in Malaysia. A total of 8 potential degraders exhibited clear zones on poly(3-hydroxybutyrate) [P(3HB)] based agar, indicating the presence of extracellular PHA depolymerase. Among the isolates, DP5 exhibited the largest clearing zone with a degradation index of 6.0. The highest degradation activity of P(3HB) was also observed with depolymerase enzyme of DP5 in mineral salt medium containing P(3HB). Based on biochemical characterization and 16S rRNA cloning and sequencing, isolate DP5 was found to belong to the genus Acidovorax and subsequently named as Acidovorax sp. DP5. The highest extracellular depolymerase enzyme activity was achieved when 0.25% (w/v) of P(3HB) and 1 g/L of urea were used as carbon and nitrogen source, respectively, in the culture media. The most suitable assay condition of the depolymerase enzyme in response to pH and temperature was tested. The depolymerase produced by strain Acidovorax sp. DP5 showed high percentage of degradation with P(3HB) films in an alkaline condition with pH 9 and at a temperature of 40°C

Elucidation of antimicrobial silver sulfadiazine (SSD) blend/poly(3-hydroxybutyrate-co-4-hydroxybutyrate) immobilised with collagen peptide as potential biomaterial

10.3390/polym12122979Polymers12121-1

Designing Novel Interfaces via Surface Functionalization of Short-Chain-Length Polyhydroxyalkanoates

Polyhydroxyalkanoates (PHA), a microbial plastic has emerged as promising biomaterial owing to the broad range of mechanical properties. However, some studies revealed that PHA is hydrophobic and has no recognition site for cell attachment and this is often a limitation in tissue engineering aspects. Owing to this, the polymer is tailored accordingly in order to enhance the biocompatibility in vivo as well as to suit the intended application. Thus far, these surface modifications have led to PHA being widely used in various biomedical and pharmaceutical applications such as cardiac patches, wound management, nerve, bone, and cartilage repair. This review addresses the surface modification on biomedical applications focusing on short-chain-length PHA such as poly(3-hydroxybutyrate) [P(3HB)], poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV)]