Evaluation of polyvinyl alcohol composite membranes containing collagen and bone particles

Composite biomaterials provide alternative materials that improve on the properties of the individual components and can be used to replace or restore damaged or diseased tissues. Typically, a composite biomaterial consists of a matrix, often a polymer, with one or more fillers that can be made up of particles, sheets or fibres. The polymer matrix can be chosen from a wide range of compositions and can be fabricated easily and rapidly into complex shapes and structures. In the present study we have examined three size fractions of collagen-containing particles embedded at up to 60% w/w in a poly(vinyl alcohol) (PVA) matrix. The particles used were bone particles, which are a mineral-collagen composite and demineralised bone, which gives naturally cross-linked collagen particles. SEM showed well dispersed particles in the PVA matrix for all concentrations and sizes of particles, with FTIR suggesting collagen to PVA hydrogen bonding. Tg of membranes shifted to a slightly lower temperature with increasing collagen content, along with a minor amount of melting point depression. The modulus and tensile strength of membranes were improved with the addition of both particles up to 10 wt%, and were clearly strengthened by the addition, although this effect decreased with higher collagen loadings. Elongation at break decreased with collagen content. Cell adhesion to the membranes was observed associated with the collagen particles, indicating a lack of cytotoxicity

Substrate-Independent Method for Growing and Modulating the Density of Polymer Brushes from Surfaces by ATRP

We describe a method for grafting PEG-based polymer chains of variable surface density using a substrate independent approach, allowing grafting from virtually any material substrate. The approach relies upon initial coupling of a macroinitiator to plasma polymer treated surfaces. The macroinitiator is a novel random terpolymer containing ATRP initiator residues, strongly negatively charged groups, and carboxylic acid moieties that facilitate covalent surface anchoring. Surface-initiated ATRP (SI-ATRP) using polyethylene glycol methyl ether methacrylate (PEGMA) at different concentrations led to grafted surfaces of controlled thickness in either the “brush” or “mushroom” morphology, which was controlled by the abundance of initiator residues in the macroinitiator. Grafted polymer layer structure was investigated via direct interaction force measurements using colloid probe atomic force microscopy (AFM). Equilibrium, hydrated graft layer thicknesses inferred from the highly repulsive AFM force data suggest that the polymer brush graft layer contained polymer chains which were fully stretched. Since the degree of stretching resulted in layer thicknesses approaching the polymer contour length, the polymer brushes studied must be very close to maximum graft density. Grafted layers where the polymer molecules were in the mushroom regime resulted in much thinner layers but the chains had greater chain entropic freedom as indicated by strongly attractive bridging interactions between tethered chains and the silica colloid probe. Use of this experimental methodology would be suitable for preparing grafted polymer layers of a preferred density free from substrate-specific linking chemistries

Substrate-Independent Method for Growing and Modulating the Density of Polymer Brushes from Surfaces by ATRP

We describe a method for grafting PEG-based polymer chains of variable surface density using a substrate independent approach, allowing grafting from virtually any material substrate. The approach relies upon initial coupling of a macroinitiator to plasma polymer treated surfaces. The macroinitiator is a novel random terpolymer containing ATRP initiator residues, strongly negatively charged groups, and carboxylic acid moieties that facilitate covalent surface anchoring. Surface-initiated ATRP (SI-ATRP) using polyethylene glycol methyl ether methacrylate (PEGMA) at different concentrations led to grafted surfaces of controlled thickness in either the “brush” or “mushroom” morphology, which was controlled by the abundance of initiator residues in the macroinitiator. Grafted polymer layer structure was investigated via direct interaction force measurements using colloid probe atomic force microscopy (AFM). Equilibrium, hydrated graft layer thicknesses inferred from the highly repulsive AFM force data suggest that the polymer brush graft layer contained polymer chains which were fully stretched. Since the degree of stretching resulted in layer thicknesses approaching the polymer contour length, the polymer brushes studied must be very close to maximum graft density. Grafted layers where the polymer molecules were in the mushroom regime resulted in much thinner layers but the chains had greater chain entropic freedom as indicated by strongly attractive bridging interactions between tethered chains and the silica colloid probe. Use of this experimental methodology would be suitable for preparing grafted polymer layers of a preferred density free from substrate-specific linking chemistries

Substrate-Independent Method for Growing and Modulating the Density of Polymer Brushes from Surfaces by ATRP

We describe a method for grafting PEG-based polymer chains of variable surface density using a substrate independent approach, allowing grafting from virtually any material substrate. The approach relies upon initial coupling of a macroinitiator to plasma polymer treated surfaces. The macroinitiator is a novel random terpolymer containing ATRP initiator residues, strongly negatively charged groups, and carboxylic acid moieties that facilitate covalent surface anchoring. Surface-initiated ATRP (SI-ATRP) using polyethylene glycol methyl ether methacrylate (PEGMA) at different concentrations led to grafted surfaces of controlled thickness in either the “brush” or “mushroom” morphology, which was controlled by the abundance of initiator residues in the macroinitiator. Grafted polymer layer structure was investigated via direct interaction force measurements using colloid probe atomic force microscopy (AFM). Equilibrium, hydrated graft layer thicknesses inferred from the highly repulsive AFM force data suggest that the polymer brush graft layer contained polymer chains which were fully stretched. Since the degree of stretching resulted in layer thicknesses approaching the polymer contour length, the polymer brushes studied must be very close to maximum graft density. Grafted layers where the polymer molecules were in the mushroom regime resulted in much thinner layers but the chains had greater chain entropic freedom as indicated by strongly attractive bridging interactions between tethered chains and the silica colloid probe. Use of this experimental methodology would be suitable for preparing grafted polymer layers of a preferred density free from substrate-specific linking chemistries

Examination of basement membrane components associated with the bovine seminiferous tubule basal lamina

© CSIRO 2007Immunohistology has been used to examine the distribution of certain components of the basement membrane (BM) associated with bovine spermatogonial germ cells that are located within the seminiferous tubules. Histology was performed on testis tissue from Brahman cattle (Bos indicus) of three different age groups: pre-pubescent (4–6 months), juvenile (8–10 months) and adult (18–24 months) animals. There were no major changes in the BM composition apparent between these three age groups, except for certain lectin staining. These data suggest that the predominant collagen type IV component may have an α3 and α4 composition, although other chains, including the α5 and α6 chains, were also present. Possibly the main laminin type present was laminin 121 (α1β2γ1), although other variants were also present. Both nidogen-1 and perlecan, which are normal BM components, were also found as part of the seminiferous tubule BM. Interstitial collagens, such as type I, III and VI collagens, were found in the peritubular space, but were not part of the BM itself, although type VI collagen was most visible in the peritubular zone adjacent to the tubules. Examination of the BM with a range of lectins gave strong staining for (glcNAc)2 entities, weak positive staining for α-L-fuc, but little or no staining for α-galNAc and (glcNAc)3 at all ages, whereas staining for α-gal, β-gal(1→3)galNAc and α-man showed developmental changes.Veronica Glattauer, Helen F. Irving-Rodgers, Raymond J. Rodgers, Sally Stockwell, Alan G. Brownlee, Jerome A. Werkmeister and John A. M. Ramsha

Substrate-Independent Method for Growing and Modulating the Density of Polymer Brushes from Surfaces by ATRP

We describe a method for grafting PEG-based polymer chains of variable surface density using a substrate independent approach, allowing grafting from virtually any material substrate. The approach relies upon initial coupling of a macroinitiator to plasma polymer treated surfaces. The macroinitiator is a novel random terpolymer containing ATRP initiator residues, strongly negatively charged groups, and carboxylic acid moieties that facilitate covalent surface anchoring. Surface-initiated ATRP (SI-ATRP) using polyethylene glycol methyl ether methacrylate (PEGMA) at different concentrations led to grafted surfaces of controlled thickness in either the “brush” or “mushroom” morphology, which was controlled by the abundance of initiator residues in the macroinitiator. Grafted polymer layer structure was investigated via direct interaction force measurements using colloid probe atomic force microscopy (AFM). Equilibrium, hydrated graft layer thicknesses inferred from the highly repulsive AFM force data suggest that the polymer brush graft layer contained polymer chains which were fully stretched. Since the degree of stretching resulted in layer thicknesses approaching the polymer contour length, the polymer brushes studied must be very close to maximum graft density. Grafted layers where the polymer molecules were in the mushroom regime resulted in much thinner layers but the chains had greater chain entropic freedom as indicated by strongly attractive bridging interactions between tethered chains and the silica colloid probe. Use of this experimental methodology would be suitable for preparing grafted polymer layers of a preferred density free from substrate-specific linking chemistries

Shaping collagen for engineering hard tissues: Towards a printomics approach

Hard tissue engineering has evolved over the past decades, with multiple approaches being explored and developed. Despite the rapid development and success of advanced 3D cell culture, 3D printing technologies and material developments, a gold standard approach to engineering and regenerating hard tissue substitutes such as bone, dentin and cementum, has not yet been realised. One such strategy that differs from conventional regenerative medicine approach of other tissues, is the in vitro mineralisation of collagen templates in the absence of cells. Collagen is the most abundant protein within the human body and forms the basis of all hard tissues. Once mineralised, collagen provides important support and protection to humans, for example in the case of bone tissue. Multiple in vitro fabrication strategies and mineralisation approaches have been developed and their success in facilitating mineral deposition on collagen to achieve bone-like scaffolds evaluated. Critical to the success of such fabrication and biomineralisation approaches is the collagen template, and its chemical composition, organisation, and density. The key factors that influence such properties are the collagen processing and fabrication techniques utilised to create the template, and the mineralisation strategy employed to deposit mineral on and throughout the templates. However, despite its importance, relatively little attention has been placed on these two critical factors. Here, we critically examine the processing, fabrication and mineralisation strategies that have been used to mineralise collagen templates, and offer insights and perspectives on the most promising strategies for creating mineralised collagen scaffolds. Statement of significance: In this review, we highlight the critical need to fabricate collagen templates with advanced processing techniques, in a manner that achieves biomimicry of the hierarchical collagen structure, prior to utilising in vitro mineralisation strategies. To this end, we focus on the initial collagen that is selected, the extraction techniques used and the native fibril forming potential retained to create reconstituted collagen scaffolds. This review synthesises current best practises in material sourcing, processing, mineralisation strategies and fabrication techniques, and offers insights into how these can best be exploited in future studies to successfully mineralise collagen templates