Swee Hin Teoh

Fused deposition modeling of novel scaffold architectures for tissue engineering applications.

Fused deposition modeling, a rapid prototyping technology, was used to produce novel scaffolds with honeycomb-like pattern, fully interconnected channel network, and controllable porosity and channel size. A bioresorbable polymer poly(epsilon-caprolactone) (PCL) was developed as a filament modeling material to produce porous scaffolds, made of layers of directionally aligned microfilaments, using this computer-controlled extrusion and deposition process. The PCL scaffolds were produced with a range of channel size 160-700 microm, filament diameter 260-370 microm and porosity 48-77%, and regular geometrical honeycomb pores, depending on the processing parameters. The scaffolds of different porosity also exhibited a pattern of compressive stress-strain behavior characteristic of porous solids under such loading. The compressive stiffness ranged from 4 to 77 MPa, yield strength from 0.4 to 3.6 MPa and yield strain from 4% to 28%. Analysis of the measured data shows a high correlation between the scaffold porosity and the compressive properties based on a power-law relationship.

Mechanical properties and cell cultural response of polycaprolactone scaffolds designed and fabricated via fused deposition modeling.

A number of different processing techniques have been developed to design and fabricate three-dimensional (3D) scaffolds for tissue-engineering applications. The imperfection of the current techniques has encouraged the use of a rapid prototyping technology known as fused deposition modeling (FDM). Our results show that FDM allows the design and fabrication of highly reproducible bioresorbable 3D scaffolds with a fully interconnected pore network. The mechanical properties and in vitro biocompatibility of polycaprolactone scaffolds with a porosity of 61 +/- 1% and two matrix architectures were studied. The honeycomb-like pores had a size falling within the range of 360 x 430 x 620 microm. The scaffolds with a 0/60/120 degrees lay-down pattern had a compressive stiffness and a 1% offset yield strength in air of 41.9 +/- 3.5 and 3.1 +/- 0.1 MPa, respectively, and a compressive stiffness and a 1% offset yield strength in simulated physiological conditions (a saline solution at 37 degrees C) of 29.4 +/- 4.0 and 2.3 +/- 0.2 MPa, respectively. In comparison, the scaffolds with a 0/72/144/36/108 degrees lay-down pattern had a compressive stiffness and a 1% offset yield strength in air of 20.2 +/- 1.7 and 2.4 +/- 0.1 MPa, respectively, and a compressive stiffness and a 1% offset yield strength in simulated physiological conditions (a saline solution at 37 degrees C) of 21.5 +/- 2.9 and 2.0 +/- 0.2 MPa, respectively. Statistical analysis confirmed that the five-angle scaffolds had significantly lower stiffness and 1% offset yield strengths under compression loading than those with a three-angle pattern under both testing conditions (p < or = 0.05). The obtained stress-strain curves for both scaffold architectures demonstrate the typical behavior of a honeycomb structure undergoing deformation. In vitro studies were conducted with primary human fibroblasts and periosteal cells. Light, environmental scanning electron, and confocal laser microscopy as well as immunohistochemistry showed cell proliferation and extracellular matrix production on the polycaprolactone surface in the 1st culturing week. Over a period of 3-4 weeks in a culture, the fully interconnected scaffold architecture was completely 3D-filled by cellular tissue. Our cell culture study shows that fibroblasts and osteoblast-like cells can proliferate, differentiate, and produce a cellular tissue in an entirely interconnected 3D polycaprolactone matrix.

Scaffold development using 3D printing with a starch-based polymer

Rapid prototyping (RP) techniques have been utilised by tissue engineers to produce three-dimensional (3D) porous scaffolds. RP technologies allow the design and fabrication of complex scaffold geometries with a fully interconnected pore network. Three-dimensional printing (3DP) technique was used to fabricate scaffolds with a novel micro- and macro-architecture. In this study, a unique blend of starch-based polymer powders (cornstarch, dextran and gelatin) was developed for the 3DP process. Cylindrical scaffolds of five different designs were fabricated and post-processed to enhance the mechanical and chemical properties. The scaffold properties were characterised by scanning electron microscopy (SEM), differential scanning calorimetry (DSC), porosity analysis and compression tests.

Knitted poly-lactide-co-glycolide scaffold loaded with bone marrow stromal cells in repair and regeneration of rabbit Achilles tendon.

The objectives of this study were to evaluate the morphology and biomechanical function of Achilles tendons regenerated using knitted poly-lactide-co-glycolide (PLGA) loaded with bone marrow stromal cells (bMSCs). The animal model used was that of an adult female New Zealand White rabbit with a 10-mm gap defect of the Achilles tendon. In group I, 19 hind legs with the created defects were treated with allogeneic bMSCs seeded on knitted PLGA scaffold. In group II, the Achilles tendon defects in 19 hind legs were repaired using the knitted PLGA scaffold alone, and in group III, 6 hind legs were used as normal control. The tendon-implant constructs of groups I and II were evaluated postoperatively at 2, 4, 8, and 12 weeks using macroscopic, histological, and immunohistochemical techniques. In addition, specimens from group I (n = 7), group II (n = 7), and group III (n = 6) were harvested for biomechanical test 12 weeks after surgery. Postoperatively, at 2 and 4 weeks, the histology of group I specimens exhibited a higher rate of tissue formation and remodeling as compared with group II, whereas at 8 and 12 weeks postoperation, the histology of both group I and group II was similar to that of native tendon tissue. The wound sites of group I healed well and there was no apparent lymphocyte infiltration. Immunohistochemical analysis showed that the regenerated tendons were composed of collagen types I and type III fibers. The tensile stiffness and modulus of group I were 87 and 62.6% of normal tendon, respectively, whereas those of group II were about 56.4 and 52.9% of normal tendon, respectively. These results suggest that the knitted PLGA biodegradable scaffold loaded with allogeneic bone marrow stromal cells has the potential to regenerate and repair gap defect of Achilles tendon and to effectively restore structure and function.

Evaluation of polycaprolactone scaffold degradation for 6 months in vitro and in vivo

The use of polycaprolactone (PCL) as a biomaterial, especially in the fields of drug delivery and tissue engineering, has enjoyed significant growth. Understanding how such a device or scaffold eventually degrades in vivo is paramount as the defect site regenerates and remodels. Degradation studies of three-dimensional PCL and PCL-based composite scaffolds were conducted in vitro (in phosphate buffered saline) and in vivo (rabbit model). Results up to 6 months are reported. All samples recorded virtually no molecular weight changes after 6 months, with a maximum mass loss of only about 7% from the PCL-composite scaffolds degraded in vivo, and a minimum of 1% from PCL scaffolds. Overall, crystallinity increased slightly because of the effects of polymer recrystallization. This was also a contributory factor for the observed stiffness increment in some of the samples, while only the PCL-composite scaffold registered a decrease. Histological examination of the in vivo samples revealed good biocompatibility, with no adverse host tissue reactions up to 6 months. Preliminary results of medical-grade PCL scaffolds, which were implanted for 2 years in a critical-sized rabbit calvarial defect site, are also reported here and support our scaffold design goal for gradual and late molecular weight decreases combined with excellent long-term biocompatibility and bone regeneration.

Fatigue of biomaterials: a review

Fatigue fracture and wear have been identified as some of the major problems associated with implant failure of medical devices. The actual in vivo mechanisms are complex and involve the hostile body environment. The response of the host tissue to wear debris is a real issue. Fatigue-wear corrosion and environmental stress cracking are common. Although fatigue fracture and wear are frequently reported in orthopaedic applications such as hip joint prostheses, they can be fatal in mechanical heart valves. While it is not possible to avoid failure, recent work has focused on predictive tools to enable more accurate prediction so as to avoid catastrophic failure in vivo. This paper presents an overview of fatigue fracture problems in metallic, polymeric and ceramic implant materials, looks at some recent techniques of testing and discusses the future development of fracture and wear resistant biomaterials.

Superior osteogenic capacity for bone tissue engineering of fetal compared with perinatal and adult mesenchymal stem cells

Mesenchymal stem cells (MSCs) from human adult bone marrow (haMSCs) represent a promising source for bone tissue engineering. However, their low frequencies and limited proliferation restrict their clinical utility. Alternative postnatal, perinatal, and fetal sources of MSCs appear to have different osteogenic capacities, but have not been systematically compared with haMSCs. We investigated the proliferative and osteogenic potential of MSCs from human fetal bone marrow (hfMSCs), human umbilical cord (hUCMSCs), and human adult adipose tissue (hATMSCs), and haMSCs, both in monolayer cultures and after loading into three‐dimensional polycaprolactone‐tricalcium‐phosphate scaffolds.Although all MSCs had comparable immunophenotypes, only hfMSCs and hUCMSCs were positive for the embryonic pluripotency markers Oct‐4 and Nanog. hfMSCs expressed the lowest HLA‐I level (55% versus 95%–99%) and the highest Stro‐1 level (51% versus 10%–27%), and had the greatest colony‐forming unit–fibroblast capacity (1.6×–2.0×; p < .01) and fastest doubling time (32 versus 54–111 hours; p < .01). hfMSCs had the greatest osteogenic capacity, as assessed by von‐Kossa staining, alkaline phosphatase activity (5.1×–12.4×; p < .01), calcium deposition (1.6×–2.7× in monolayer and 1.6×–5.0× in scaffold culture; p < .01), calcium visualized on micro‐computed tomography (3.9×17.6×; p < .01) and scanning electron microscopy, and osteogenic gene induction. Two months after implantation of cellular scaffolds in immunodeficient mice, hfMSCs resulted in the most robust mineralization (1.8×–13.3×; p < .01).The ontological and anatomical origins of MSCs have profound influences on the proliferative and osteogenic capacity of MSCs. hfMSCs had the most proliferative and osteogenic capacity of the MSC sources, as well as being the least immunogenic, suggesting they are superior candidates for bone tissue engineering. STEM CELLS 2009;27:126–137

Dynamics of in vitro polymer degradation of polycaprolactone-based scaffolds: accelerated versus simulated physiological conditions

The increasing use of biodegradable devices in tissue engineering and regenerative medicine means it is essential to study and understand their degradation behaviour. Accelerated degradation systems aim to achieve similar degradation profiles within a shorter period of time, compared with standard conditions. However, these conditions only partially mimic the actual situation, and subsequent analyses and derived mechanisms must be treated with caution and should always be supported by actual long-term degradation data obtained under physiological conditions. Our studies revealed that polycaprolactone (PCL) and PCL-composite scaffolds degrade very differently under these different degradation conditions, whilst still undergoing hydrolysis. Molecular weight and mass loss results differ due to the different degradation pathways followed (surface degradation pathway for accelerated conditions and bulk degradation pathway for simulated physiological conditions). Crystallinity studies revealed similar patterns of recrystallization dynamics, and mechanical data indicated that the scaffolds retained their functional stability, in both instances, over the course of degradation. Ultimately, polymer degradation was shown to be chiefly governed by molecular weight, crystallinity susceptibility to hydrolysis and device architecture considerations whilst maintaining its thermodynamic equilibrium.

Review: development of clinically relevant scaffolds for vascularised bone tissue engineering.

Clinical translation of scaffold-based bone tissue engineering (BTE) therapy still faces many challenges despite intense investigations and advancement over the years. To address these clinical barriers, it is important to analyse the current technical challenges in constructing a clinically relevant scaffold and subsequent clinical issues relating to bone repair. This review highlights the key challenges hampering widespread clinical translation of scaffold-based vascularised BTE, with a focus on the repair of large non-union defects. The main limitations of current scaffolds include the lack of sufficient vascularisation, insufficient mechanical strength as well as issues relating to the osseointegration of the bioresorbable scaffold and bone infection management. Critical insights on the current trends of scaffold technologies and future directions for advancing next-generation BTE scaffolds into the clinical realm are discussed. Considerations concerning regulatory approval and the route towards commercialisation of the scaffolds for widespread clinical utility will also be introduced.

Fabrication of 3D chitosan–hydroxyapatite scaffolds using a robotic dispensing system

Abstract A new robotic desktop rapid prototyping (RP) system was designed to fabricate scaffolds for tissue engineering applications. The experimental setup consists of a computer-guided desktop robot and a one-component pneumatic dispenser. The dispensing material (chitosan and chitosan–hydroxyapatite (HA) dissolved in acetic acid) was stored in a 30-ml barrel and forced out through a small Teflon-lined nozzle into a dispensing medium (sodium hydroxide–ethanol in ratio of 7:3). Layer-by-layer, the chitosan was fabricated with a preprogramed lay-down pattern. Neutralization of the chitosan forms a gel-like precipitate, and the hydrostatic pressure in the sodium hydroxide (NaOH) solution keeps the cuboid scaffold in shape. Comparison of the freeze-dried scaffold to the wet one showed linear and volumetric shrinkage of about 31% and 62%, respectively. A good attachment between layers allowed the chitosan matrix to form a fully interconnected channel architecture. Results of in vitro cell culture studies revealed the scaffold biocompatibility. The results of this preliminary study using the rapid prototyping robotic dispensing (RPBOD) system demonstrated its potential in fabricating three-dimensional (3D) scaffolds with regular and reproducible macropore architecture.