Anatomy Articles

Examination of osteoarthritis and subchondral bone alterations within the stifle joint of an ovariectomised ovine model.

Jane C. Holland et al. — Thu, 23 May 2013 07:31:48 PDT

The exact relationship between osteoporosis and osteoarthritis is still a matter for debate for many. The ovariectomised ewe is frequently used as a model for osteoporosis, resulting in significant alterations in bone morphometry and turnover in both trabecular and subchondral bone after 1 year. This study examines whether ovariectomy has any impact on development of osteoarthritis within the ovine stifle joint at the same time point. In addition, we investigate whether there are any significant correlations present between articular cartilage degeneration and alterations in microstructural parameters or turnover rates in the underlying bone. Twenty-two sheep were examined in this study; 10 of the sheep underwent ovariectomy and 12 were kept as controls. Five distinctive fluorochrome dyes were administered intravenously at 12-week intervals to both groups, to label sites of bone turnover. All animals were then sacrificed 12 months postoperatively. Although most specimens showed some evidence of osteoarthritis, no measurable difference between the two study groups was detected. Osteoarthritis was associated with a thinning of the subchondral plate, specifically the subchondral cortical bone; however, whereas previous studies have suggested a link between trabecular thinning and osteoarthritis, this was not confirmed. No correlation was found between osteoarthritis and bone turnover rates of either the subchondral trabecular bone or bone plate. In conclusion, despite the fact that ovariectomy results in marked morphological and structural changes in the ovine stifle joint at 1-year postoperatively, no evidence was found to suggest that it plays a direct role in the aetiology of osteoarthritis.

Subchondral osteopenia and accelerated bone remodelling post-ovariectomy - a possible mechanism for subchondral microfractures in the aetiology of spontaneous osteonecrosis of the knee?

Jane C. Holland et al. — Tue, 22 Jan 2013 08:20:38 PST

Osteopenia and subchondral microfractures are implicated in the aetiology of spontaneous osteonecrosis of the knee. The ovine tibia shows significant alterations of the trabecular architecture within the subchondral bone of the medial tibial plateau post-ovariectomy, including reduced trabecular bone volume fraction. We hypothesise that accelerated subchondral bone resorption may also play a role in increasing microfracture risk at this site. 23 sheep were examined in this study; 10 of the sheep underwent ovariectomy (OVX), while the remainder (n=13) were kept as controls (CON). Five fluorochrome dyes were administered intravenously at 12 week intervals via the jugular vein to both groups, to label sites of bone turnover. These animals were then sacrificed at 12 months post-operatively. Bone turnover was significantly increased in the OVX group in both trabecular bone (2.024 vs. 1.047, p = 0.05) and within the subchondral bone plate (4.68 vs. 0.69 # / mm2; p < 0.001). In addition to the classically-described turnover visible along trabecular surfaces, we also found visual evidence of intra-trabecular osteonal remodelling. In conclusion, this study shows significant alterations in bone turnover in both trabecular bone and within the subchondral bone plate at one-year post-ovariectomy. Remodelling of trabecular bone was due to both classically described hemi-osteonal and intra-trabecular osteonal remodelling. The presence of both localised osteopenia and accelerated bone remodelling within the medial tibial plateau provide a possible mechanism for subchondral microfractures in the aetiology of spontaneous osteonecrosis of the knee. Further utilisation of the ovariectomised ewe may be useful for further study in this field.

Temporal changes in bone composition, architecture, and strength following estrogen deficiency in osteoporosis.

Orlaith Brennan et al. — Tue, 04 Dec 2012 02:08:38 PST

Using an ovariectomized (OVX) ovine model, we provide an analysis of the timing of changes in bone following estrogen deficiency. The expression of genes known to regulate osteoclastogenesis, matrix production, and mineralization, as measured by real-time RT-PCR, was significantly increased by 12 months; and increased expression was maintained through to 31 months post-OVX compared to controls. FTIR spectroscopy confirmed that mineralized crystals were less mature than in controls 12 months post-OVX and were even less so by 31 months. The mineral-to-matrix ratio was significantly reduced by 31 months, while the ratio of mature to immature collagen cross-linking was initially increased at 12 months and subsequently reduced at 31 months post-OVX. In contrast, trabecular number, thickness, and separation were unchanged at 12 months. Significant reductions in trabecular number and thickness and a significant increase in trabecular separation were observed 31 months after OVX. Most notably perhaps these combined changes led to a significant reduction in the compressive strength of trabecular bone after 31 months. The results indicate that there is an initial increase in bone turnover, which is accompanied by a change in bone composition. This is followed by a continued increase in bone resorption and relative reduction in bone formation, leading to deterioration in bone microarchitecture. Ultimately, these cumulative changes led to a significant reduction in the compressive strength of bones following 31 months of estrogen deficiency. These findings provide important insight into the time sequence of changes during osteoporosis.

Mechanical characterization of a customized decellularized scaffold for vascular tissue engineering.

W S. Sheridan et al. — Tue, 27 Nov 2012 07:01:34 PST

Several challenges persist when attempting to utilize decellularized tissue as a scaffold for vascular tissue engineering. Namely: poor cell infiltration/migration, excessive culture times associated with repopulating the scaffolds, and the achievement of a quiescent medial layer. In an attempt to create an optimum vascular scaffold, we customized the properties of decellularized porcine carotid arteries by: (i) creating cavities within the medial layer to allow direct injection of cells, and (ii) controlling the amount of collagen digestion to increase the porosity. Histological examination of our customized scaffold revealed a highly porous tissue structure containing consistent medial cavities running longitudinally through the porous scaffold wall. Mechanical testing of the customized scaffold showed that our minimal localized disruption to the ECM does not have a detrimental effect on the bulk mechanical response of the tissue. The results demonstrate that an increased stiffness and reduced distensibility occurs after decellularization when compared to the native tissue, however post scaffold customization we can revert the scaffold tensile properties back to that of the native tissue. This most noteworthy result occurs in the elastin dominant phase of the tensile response of the scaffold, indicating that no disruption has occurred to the elastin network by our decellularization and customization techniques. Additionally, the bulk seeding potential of the customized scaffold was demonstrated by direct injection of human smooth muscle cells through the medial cavities. The optimum cell dispersion was observed in the highest porosity scaffold, with large cell numbers retained within the medial layer after 24 h static culture. In summary, this study presents a novel customized decellularized vascular scaffold that has the capability of bulk seeding the media, and in tandem to this method, the porosity of the scaffold has been increased without compromising the mechanical integrity.

Distribution of microcrack lengths in bone in vivo and in vitro.

Gerardo Presbitero et al. — Tue, 27 Nov 2012 06:40:52 PST

It is well known that bone contains small cracks; in vivo these microcracks are constantly growing and being repaired. Too rapid crack growth leads to stress fractures or fragility fractures. In vitro, changes occur in this population of microcracks when subjected to cyclic loading up to and including failure. Normally, the only parameters reported from such investigations are the number density of cracks and their average length. In the present work we examined the microcrack population in more detail. We analysed ten different sets of experimental data including in vivo and in vitro microcracks, plus two theoretical simulations. We showed for the first time that the distribution of crack lengths can be described using the two-parameter Weibull equation. The values of the two constants in the equation varied depending on bone type/species and showed consistent trends during in vitro testing. This is the most detailed study to be conducted on microcrack populations in bone; the results will be useful in future studies including the development of theoretical models and computer simulations of bone damage and failure.

Visualising feasible operating ranges within tissue engineering systems using a "windows of operation" approach: A perfusion-scaffold bioreactor case study.

Ryan J. McCoy et al. — Wed, 30 May 2012 06:47:39 PDT

Tissue engineering approaches to developing functional substitutes are often highly complex, multivariate systems where many aspects of the biomaterials, bio-regulatory factors or cell sources may be controlled in an effort to enhance tissue formation. Furthermore, success is based on multiple performance criteria reflecting both the quantity and quality of the tissue produced. Managing the trade-offs between different performance criteria is a challenge. A "windows of operation" tool that graphically represents feasible operating spaces to achieve user-defined levels of performance has previously been described by researchers in the bio-processing industry. This paper demonstrates the value of "windows of operation" to the tissue engineering field using a perfusion-scaffold bioreactor system as a case study. In our laboratory, perfusion bioreactor systems are utilised in the context of bone tissue engineering to enhance the osteogenic differentiation of cell-seeded scaffolds. A key challenge of such perfusion bioreactor systems is to maximise the induction of osteogenesis but minimise cell detachment from the scaffold. Two key operating variables that influence these performance criteria are the mean scaffold pore size and flow-rate. Using cyclooxygenase-2 and osteopontin gene expression levels as surrogate indicators of osteogenesis, we employed the "windows of operation" methodology to rapidly identify feasible operating ranges for the mean scaffold pore size and flow-rate that achieved user-defined levels of performance for cell detachment and differentiation. Incorporation of such tools into the tissue engineer's armoury will hopefully yield a greater understanding of the highly complex systems used and help aid decision making in future translation of products from the bench top to the market place. Biotechnol. Bioeng. © 2012 Wiley Periodicals, Inc.

Influence of flow rate and scaffold pore size on cell behavior during mechanical stimulation in a flow perfusion bioreactor.

Ryan J. McCoy et al. — Fri, 17 Feb 2012 02:32:25 PST

Mechanically stimulating cell-seeded scaffolds by flow-perfusion is one approach utilized for developing clinically applicable bone graft substitutes. A key challenge is determining the magnitude of stimuli to apply that enhances cell differentiation but minimizes cell detachment from the scaffold. In this study, we employed a combined computational modeling and experimental approach to examine how the scaffold mean pore size influences cell attachment morphology and subsequently impacts upon cell deformation and detachment when subjected to fluid-flow. Cell detachment from osteoblast-seeded collagen-GAG scaffolds was evaluated experimentally across a range of scaffold pore sizes subjected to different flow rates and exposure times in a perfusion bioreactor. Cell detachment was found to be proportional to flow rate and inversely proportional to pore size. Using this data, a theoretical model was derived that accurately predicted cell detachment as a function of mean shear stress, mean pore size, and time. Computational modeling of cell deformation in response to fluid flow showed the percentage of cells exceeding a critical threshold of deformation correlated with cell detachment experimentally and the majority of these cells were of a bridging morphology (cells stretched across pores). These findings will help researchers optimize the mean pore size of scaffolds and perfusion bioreactor operating conditions to manage cell detachment when mechanically simulating cells via flow perfusion. Biotechnol. Bioeng. © 2012 Wiley Periodicals, Inc.

Temporal and spatial changes in cartilage-matrix-specific gene expression in mesenchymal stem cells in response to dynamic compression.

Matthew G. Haugh et al. — Fri, 10 Feb 2012 10:06:41 PST

Various forms of mechanical stimulation have been shown to enhance chondrogenesis of mesenchymal stem cells (MSCs). However, the response of MSCs undergoing chondrogenesis to such signals has been shown to depend on the temporal application of loading. The objective of this study was to determine the effect of dynamic compression on cartilage-matrix-specific gene expression and to relate this response to the local biochemical environment and cell phenotype at the time of loading. At 0, 7, 14, and 21 days extracellular matrix (ECM) deposition within MSC-seeded agarose hydrogels due to transforming growth factor-β3 stimulation was determined biochemically and histologically, and then reverse transcription-polymerase chain reaction was used to examine the effects of dynamic compression on cartilage-matrix-specific gene expression. The results of these experiments show that the local environment in the core of the constructs is more favorable for chondrogenesis in comparison to the annulus, as evident from both ECM synthesis and gene expression. Additionally, we found that the response of the cells to mechanical stimulus varied with both the spatial region within the constructs and the temporal application of loading. Dynamic compression applied at day 21 was found to enhance levels of cartilage matrix gene expression following a peak in expression levels at day 14 in free swelling constructs, suggesting that mechanical signals play a key role in the maintenance of a chondrogenic phenotype. The application of mechanical stimulus to enhance cartilage ECM synthesis may be an important tool in regenerative medicine-based cartilage repair. The results of this study suggest that a chondrogenic phenotype and/or a well-developed pericellular matrix must first be established before dynamic compression can have a positive effect on cartilage-matrix-specific gene expression.

Addition of hyaluronic acid improves cellular infiltration and promotes early-stage chondrogenesis in a collagen-based scaffold for cartilage tissue engineering.

Amos Matsiko et al. — Fri, 10 Feb 2012 09:58:17 PST

The response of mesenchymal stem cells (MSCs) to a matrix largely depends on the composition as well as the extrinsic mechanical and morphological properties of the substrate to which they adhere to. Collagen-glycosaminoglycan (CG) scaffolds have been extensively used in a range of tissue engineering applications with great success. This is due in part to the presence of the glycosaminoglycans (GAGs) in complementing the biofunctionality of collagen. In this context, the overall goal of this study was to investigate the effect of two GAG types: chondroitin sulphate (CS) and hyaluronic acid (HyA) on the mechanical and morphological characteristics of collagen-based scaffolds and subsequently on the differentiation of rat MSCs in vitro. Morphological characterisation revealed that the incorporation of HyA resulted in a significant reduction in scaffold mean pore size (93.9 μm) relative to collagen-CS (CCS) scaffolds (136.2 μm). In addition, the collagen-HyA (CHyA) scaffolds exhibited greater levels of MSC infiltration in comparison to the CCS scaffolds. Moreover, these CHyA scaffolds showed significant acceleration of early stage gene expression of SOX-9 (approximately 60-fold higher, p<0.01) and collagen type II (approximately 35-fold higher, p<0.01) as well as cartilage matrix production (7-fold higher sGAG content) in comparison to CCS scaffolds by day 14. Combining their ability to stimulate MSC migration and chondrogenesis in vitro, these CHyA scaffolds show great potential as appropriate matrices for promoting cartilage tissue repair.

Evaluation of the ability of collagen-glycosaminoglycan scaffolds with or without mesenchymal stem cells to heal bone defects in Wistar rats.

Mohamed Alhag et al. — Fri, 04 Nov 2011 04:49:55 PDT

PURPOSE: The aim of this experiment was to examine the capacity of collagen-glycosaminoglycan scaffolds, with or without mesenchymal stem cells, to satisfactorily repair a 5-mm rat calvarial defect. METHODS: Fifty-five Wistar rats were used in the study. The defects were either left empty to serve as controls (n = 7) or filled with cell-free scaffolds (n = 11), cell-seeded scaffolds that were pre-cultured in standard culture medium (n = 13), cell-seeded scaffolds that were pre-cultured in osteoinductive factor-supplemented medium (n = 12) or particulate autogenous bone (n = 12). The animals were sacrificed at 12 weeks after surgery, and specimens were prepared for histomorphometric analysis. The linear bone healing and the bone area within the defect were measured. RESULTS: Comparable results were obtained using cell-free collagen-glycosaminoglycan scaffolds and autogenous bone both in terms of linear bone healing (P < 0.986) and area of new bone (P < 0.846). While the test groups showed significantly more bone formation compared to the empty defect control group, the linear bone healing and area of new bone within the defect were significantly lower in the cell-seeded scaffolds than in the cell-free scaffolds. The results have demonstrated that a cell-free collagen-glycosaminoglycan scaffold is capable of repairing a 5-mm rat calvarial defect as effectively as autogenous bone and that seeding the scaffold with pre-cultured mesenchymal stem cells prior to implantation offered no beneficial effect and resulted in incomplete healing of the defect. CONCLUSIONS: The results thus suggest that the scaffold has immense potential for tissue repair showing favorable osteoconductive properties, biocompatibility and degradability.

Subchondral trabecular structural changes in the proximal tibia in an ovine model of increased bone turnover.

Jane C. Holland et al. — Wed, 02 Nov 2011 07:41:10 PDT

Ovariectomized (OVX) sheep are now considered to be useful models for a variety of metabolic bone disorders. The specific aim of this study was to determine the effects of ovariectomy on the structural parameters and material density of the subchondral bone of the ovine tibial plateau as measured by microcomputed tomography (MicroCT). Twenty-three sheep were examined in this study; 10 of the sheep underwent ovariectomy (OVX), and the remainder (n=13) were kept as controls (CON). These animals were then sacrificed at 12 months post-operatively. Three-dimensional analyses were performed of osteochondral samples (15 mm deep) which were obtained from the medial tibial plateau using MicroCT. Bone volume fraction of the subchondral trabecular bone was reduced in the ovariectomized sheep as compared to control animals (0.439 vs. 0.483, P=0.038). Trabeculae were also significantly thinner in the OVX group (0.220 vs. 0.252 mm, P=0.010), with reduced connectivity density (7.947 vs. 11.524 mm(-3) , P=0.014). There was a trend towards lower numbers of individual trabeculae present in the OVX group as compared to controls, but this did not reach significance (2.817 vs. 3.288 mm(-1) , P=0.1). There was also increased trabecular separation in the OVX group, which again fell short of significance (0.426 vs. 0.387 mm, P=0.251). There was no difference in hydroxyapatite concentration (HA) between the two groups (929 vs. 932 mgHA cm(-3) , P=0.687). In conclusion, significant alterations of the trabecular architecture under the tibial plateau were observed following 12 months of oestrogen-deficiency in this ovine model. Despite these marked morphological and structural density differences, the material densities were equal in the two groups.

The Marine-derived, Multi-mineral formula, Aquamin, Enhances Mineralisation of Osteoblast Cells In Vitro.

Denise M O'Gorman et al. — Wed, 02 Nov 2011 05:02:08 PDT

Osteoporosis is a global health problem characterized by low bone mass and an increase in bone fragility. It is now well accepted that dietary factors play a central role in bone development and health. Diet that lacks adequate minerals is considered to be a risk factor for osteoporosis. The food supplement, Aquamin, is a natural, multi-mineral derived from the red algae Lithothamnion corallioides, rich in calcium, magnesium and 72 other trace minerals. The aim of this study was to evaluate the effect of Aquamin on osteoblastic behaviour and mineralisation in a pre-osteoblastic cell line. Cell number and metabolic activity were assessed using Hoescht DNA and AlamarBlue assays respectively. Osteogenic differentiation was measured using an alkaline phosphatase assay while mineralisation was determined using von Kossa and alizarin red staining. It is reported here that Aquamin promotes increased mineralisation in osteoblast cell culture. These data suggest that the nutritional supplement Aquamin plays an important role in promoting bone formation and may be useful in treating bone diseases such as osteoporosis. Copyright © 2011 John Wiley & Sons, Ltd.

IV.3. Bioreactors in tissue engineering.

Niamh A. Plunkett et al. — Fri, 17 Jun 2011 07:50:58 PDT

Osteoblast response to rest periods during bioreactor culture of collagen-glycosaminoglycan scaffolds.

Niamh A. Plunkett et al. — Fri, 17 Jun 2011 07:38:56 PDT

Flow perfusion bioreactors have been shown to enhance fluid transport and improve cell viability throughout tissue-engineered bone constructs. Furthermore, osteoblasts have been shown to be stimulated by flow during bioreactor culture, although the optimum flow regime to promote an osteogenic response has yet to be found. One problem is that bone cells lose their ability to respond to stimulation; however, mechanosensitivity can be restored by introducing resting periods between bouts of loading. The aim of this study was to analyze the effect of rest-insertion on the response of osteoblasts seeded on collagen-glycosaminoglycan scaffolds in a flow perfusion bioreactor over culture periods up to 14 days. Short-term rests of 5, 10, or 15 s and long-term rests of 7 h were incorporated into stimulation patterns. Cell distribution was enhanced in all flow groups, whereas static culture controls exhibited encapsulation. Cyclooxygenase-2 expression and prostaglandin E(2) levels increased significantly because of bioreactor culture over static controls. Osteopontin expression was significantly higher for the rest-inserted groups than the static control group or steady-flow group. These results indicate that the insertion of resting periods during flow enhances cellular distribution and osteogenic responses on collagen-glycosaminoglycan constructs cultured in a flow perfusion bioreactor.

3 hours of perfusion culture prior to 28 days of static culture, enhances osteogenesis by human cells in a collagen GAG scaffold.

Michael B. Keogh et al. — Wed, 23 Mar 2011 08:43:24 PDT

In tissue engineering bioreactors can be used to aid in the in vitro development of new tissue by providing biochemical and physical regulatory signals to cells and encouraging them to undergo differentiation and/or to produce extracellular matrix prior to in vivo implantation. This study examined the effect of short term flow perfusion bioreactor culture, prior to long term static culture, on human osteoblast cell distribution and osteogenesis within a collagen glycosaminoglycan (CG) scaffold for bone tissue engineering. Human Foetal Osteoblasts (hFOB 1.19) were seeded onto CG scaffolds and pre-cultured for 6 days. Constructs were then placed into the bioreactor and exposed to 3×1hr bouts of steady flow (1ml/min) separated by 7hrs of no flow over a 24hr period. The constructs were then cultured under static osteogenic conditions for up to 28 days. Results show that the bioreactor and static culture control groups displayed similar cell numbers and metabolic activity. Histologically however, peripheral cell-encapsulation was observed in the static controls, whereas, improved migration and homogenous cell distribution was seen in the bioreactor groups. Gene expression analysis showed that all osteogenic markers investigated displayed greater levels of expression in the bioreactor groups compared to static controls. While static groups showed increased mineral deposition; mechanical testing revealed that there was no difference in the compressive modulus between bioreactor and static groups. In conclusion, a flow perfusion bioreactor improved construct homogeneity by preventing peripheral encapsulation whilst also providing an enhanced osteogenic phenotype over static controls. © 2010 Wiley Periodicals, Inc.

Development and characterisation of a collagen nano-hydroxyapatite composite scaffold for bone tissue engineering.

Grainne M. Cunniffe et al. — Wed, 23 Mar 2011 08:27:11 PDT

Bone regeneration requires scaffolds that possess suitable mechanical and biological properties. This study sought to develop a novel collagen-nHA biocomposite scaffold via two new methods. Firstly a stable nHA suspension was produced and added to a collagen slurry (suspension method), and secondly, porous collagen scaffolds were immersed in nHA suspension after freeze-drying (immersion method). Significantly stronger constructs were produced using both methods compared to collagen only scaffolds, with a high porosity maintained (>98.9%). It was found that Coll-nHA composite scaffolds produced by the suspension method were up to 18 times stiffer than the collagen control (5.50 +/- 1.70 kPa vs. 0.30 +/- 0.09 kPa). The suspension method was also more reproducible, and the quantity of nHA incorporated could be varied with greater ease than with the immersion technique. In addition, Coll-nHA composites display excellent biological activity, demonstrating their potential as bone graft substitutes in orthopaedic regenerative medicine.

The synthesis and characterization of nanophase hydroxyapatite using a novel dispersant-aided precipitation method.

Grainne M. Cunniffe et al. — Wed, 23 Mar 2011 08:13:55 PDT

The synthesis of nanophase hydroxyapatite (nHA) is of importance in the field of biomaterials and bone tissue engineering. The bioactive and osteoconductive properties of nHA are of much benefit to a wide range of biomedical applications such as producing bone tissue engineered constructs, coating medical implants, or as a carrier for plasmid DNA in gene delivery. This study aimed to develop a novel low-temperature dispersant-aided precipitation reaction to produce nHA particles (<100 >nm), which are regarded as being preferable to micron-sized agglomerates of nHA. The variables investigated and optimized include the reaction pH, the rate of reactant mixing, use of sonication, order of addition, and concentration of the primary reactants, in addition, the effect of using poly(vinyl alcohol) (PVA) surfactant and Darvan 821A® dispersing agent during the reaction was also examined. It was found that by fine-tuning the synthesis parameters and incorporating the dispersing agent, monodisperse, phase-pure nano-sized particles under 100 nm were attained, suitable for clinical applications in bone regeneration.

Crosslinking and Mechanical Properties Significantly Influence Cell Attachment, Proliferation, and Migration Within Collagen Glycosaminoglycan Scaffolds.

Matthew G. Haugh et al. — Wed, 23 Mar 2011 07:44:02 PDT

Crosslinking and the resultant changes in mechanical properties have been shown to influence cellular activity within collagen biomaterials. With this in mind, we sought to determine the effects of crosslinking on both the compressive modulus of collagen-glycosaminoglycan scaffolds and the activity of osteoblasts seeded within them. Dehydrothermal, 1-ethyl-3-3-dimethyl aminopropyl carbodiimide and glutaraldehyde crosslinking treatments were first investigated for their effect on the compressive modulus of the scaffolds. After this, the most promising treatments were used to study the effects of crosslinking on cellular attachment, proliferation, and infiltration. Our experiments have demonstrated that a wide range of scaffold compressive moduli can be attained by varying the parameters of the crosslinking treatments. 1-Ethyl-3-3-dimethyl aminopropyl carbodiimide and glutaraldehyde treatments produced the stiffest scaffolds (fourfold increase when compared to dehydrothermal crosslinking). When cells were seeded onto the scaffolds, the stiffest scaffolds also showed increased cell number and enhanced cellular distribution when compared to the other groups. Taken together, these results indicate that crosslinking can be used to produce collagen-glycosaminoglycan scaffolds with a range of compressive moduli, and that increased stiffness enhances cellular activity within the scaffolds.

Novel freeze-drying methods to produce a range of collagen-glycosaminoglycan scaffolds with tailored mean pore sizes.

Matthew G. Haugh et al. — Wed, 23 Mar 2011 05:39:51 PDT

The pore structure of three-dimensional scaffolds used in tissue engineering has been shown to significantly influence cellular activity. As the optimal pore size is dependant on the specifics of the tissue engineering application, the ability to alter the pore size over a wide range is essential for a particular scaffold to be suitable for multiple applications. With this in mind, the aim of this study was to develop methodologies to produce a range of collagen-glycosaminoglycan (CG) scaffolds with tailored mean pore sizes. The pore size of CG scaffolds is established during the freeze-drying fabrication process. In this study, freezing temperature was varied (−10 degrees C to −70 degrees C) and an annealing step was introduced to the process to determine their effects on pore size. Annealing is an additional step in the freeze-drying cycle that involves raising the temperature of the frozen suspension to increase the rate of ice crystal growth. The results show that the pore size of the scaffolds decreased as the freezing temperature was reduced. Additionally, the introduction of an annealing step during freeze-drying was found to result in a significant increase (40%) in pore size. Taken together, these results demonstrate that the methodologies developed in this study can be used to produce a range of CG scaffolds with mean pore sizes from 85 to 325 microm. This is a substantial improvement on the range of pore sizes that were possible to produce previously (96-150 microm). The methods developed in this study provide a basis for the investigation of the effects of pore size on both in vitro and in vivo performance and for the determination of the optimal pore structure for specific tissue engineering applications.

Biomaterials and scaffolds for tissue engineering

Fergal J. O'Brien — Wed, 23 Mar 2011 05:09:21 PDT

Every day thousands of surgical procedures are performed to replace or repair tissue that has been damaged through disease or trauma. The developing field of tissue engineering (TE) aims to regenerate damaged tissues by combining cells from the body with highly porous scaffold biomaterials, which act as templates for tissue regeneration, to guide the growth of new tissue. This article describes the functional requirements, and types, of materials used in developing state of the art of scaffolds for tissue engineering applications. Furthermore, it describes the challenges and where future research and direction is required in this rapidly advancing field.