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ABSTRACT: Bone grafting procedures have become common due in part to a global trend of population aging. Native bone graft is a popular choice when compared to various synthetic bone graft substitutes, owing to superior biological activity. Nonetheless, the insufficient ability of bone allograft to induce new bone formation and the insufficient remodeling of native bone grafts call for osteoinductive factors during bone repair, exemplified by recombinant human bone morphogenetic protein 2 (rhBMP2). We previously developed a modular bone morphogenetic peptide (mBMP) to address complications associated with the clinical use of rhBMP2 as a bone graft substitute. The mBMP is designed to strongly bind to hydroxyapatite, the main inorganic component of bone and teeth, and to provide pro-osteogenic properties analogous to rhBMP2. Our previous in vivo animal studies showed that mBMP bound to hydroxyapatite-coated orthopedic implants with high affinity and stimulated new bone formation. In this study, we demonstrate specific binding of mBMP to native bone grafts. The results show that mBMP binds with high affinity to both cortical and trabecular bones, and that the binding is dependent on the mBMP concentration and incubation time. Importantly, efficient mBMP binding is achieved in an ex vivo bone bioreactor where bone tissue is maintained viable for several weeks. In addition, mBMP binding can be localized with spatial control on native bone tissue via simple methods, such as dip-coating, spotting and direct writing. Taken together with the pro-osteogenic activity of mBMP established in previous bone repair models, these results suggest that mBMP may promote bone healing when coated on native bone grafts in a clinically compatible manner.
Molecular Pharmaceutics 03/2013; · 4.78 Impact Factor
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ABSTRACT: VEGF-binding peptide ligands are incorporated into hydrogel microspheres and reduce the amount of growth factor in solution. VEGF binding affinity is enhanced by creating ligands with a dimer structure. The spheres are able to knock down VEGF-mediated HUVEC growth and reduce calcium signaling. The binding interaction is reversible, allowing the spheres to be used as a VEGF delivery vehicle.
Advanced healthcare materials. 07/2012; 1(4):457-60.
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ABSTRACT: Growth factor signaling plays an essential role in regulating processes such as tissue development, maintenance, and repair. Gene expression levels, diffusion, degradation, and sequestration by extracellular matrix components all play a role in regulating the concentration of growth factors within the cellular microenvironment. Herein, we describe the synthesis and characterization of hydrogel microspheres that mimic the ability of the native extracellular matrix to reversibly bind vascular endothelial growth factor (VEGF) out of solution. A peptide ligand derived from the VEGF receptor 2 (VEGFR2) was covalently incorporated into the hydrogel microspheres in order to achieve binding affinity and specificity. In addition to being able to both bind and release VEGF in a controllable manner, the microspheres were also shown to affect human umbilical vein endothelial cell (HUVEC) proliferation. The resulting microspheres may enable new strategies to specifically upregulate or downregulate growth factor signaling in the cellular microenvironment.
Biomaterials 04/2012; 33(12):3475-84. · 7.40 Impact Factor
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ABSTRACT: β-Tricalcium phosphate (β-TCP) is an attractive ceramic for bone tissue repair because of its similar composition to bone mineral and its osteoconductivity. However, compared with other ceramics β-TCP has a rapid and uncontrolled rate of degradation. In the current study β-TCP granules were mineral coated with the aim of influencing the dissolution rate of β-TCP, and also to use the coating as a carrier for controlled release of the growth factors recombinant human vascular endothelial growth factor (rhVEGF), modular VEGF peptide (mVEGF), and modular bone morphogenetic protein 2 peptide (mBMP2). The biomineral coatings were formed by heterogeneous nucleation in aqueous solution using simulated body fluid solutions with varying concentrations of bicarbonate (HCO(3)). Our results demonstrate that we could coat β-TCP granules with mineral layers possessing different dissolution properties. The presence of a biomineral coating delays the dissolution rate of the β-TCP granules. As the carbonate (CO(3)(2-)) content in the coating was increased the dissolution rate of the coated β-TCP also increased, but remained slower than the dissolution of uncoated β-TCP. In addition, we showed sustained release of multiple growth factors, with release kinetics that were controllable by varying the identity of the growth factor or the CO(3)(2-) content in the mineral coating. Released rhVEGF induced human umbilical vein endothelial cell (HUVEC) proliferation, and mVEGF enhanced migration of mouse embryonic endothelial cells in a scratch wound healing assay, indicating that each released growth factor was biologically active.
Acta biomaterialia 12/2011; 8(3):1117-24. · 3.98 Impact Factor
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ABSTRACT: In this work, we develop and validate an automated micro-computed tomography (micro-CT) image segmentation algorithm that accurately and efficiently segments bone, calcium phosphate (CaP)-based bone scaffold, and soft tissue. The algorithm enables quantitative evaluation of bone growth in CaP scaffolds in our study that includes many samples (100+) and large data sets (900 images per sample). The use of micro-CT for such applications is otherwise limited because the similarity in X-ray attenuation for the two materials makes them indistinguishable. Destructive characterization using histological techniques and scanning electron microscopy (SEM) has been the standard for CaP scaffolds, but these methods are cumbersome, inaccurate, and yield only 2D information. The proposed algorithm exploits scaffold periodicity and combines signal analysis, edge detection, and knowledge of three-dimensional spatial relationships between bone, CaP scaffold, and soft tissue to achieve fast and accurate segmentation. Application of this algorithm can lead to a new understanding of the role of CaP and scaffold internal structure on patterns and rates of bone growth.
Computerized medical imaging and graphics: the official journal of the Computerized Medical Imaging Society 08/2011; 36(1):54-65. · 1.04 Impact Factor
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ABSTRACT: An important consideration in bone regeneration is the need for expedited neovascularization within the defect site. Formation of a vascular network is critical for cell viability and normal function leading to tissue regeneration, but spontaneous angiogenesis is too slow to yield sufficient vessel formation. In this pilot study, human umbilical cord blood (hUCB)-derived endothelial colony forming cells (ECFCs) were evaluated for in vivo vasculogenesis in the macropores of biphasic calcium phosphate (BCP)/bone morphogenetic protein-2 (BMP-2) bone tissue engineering constructs. Constructs were implanted on the abdominal wall of NOD/SCID mice for 4 weeks. This study demonstrated in vivo vasculogenesis by human ECFCs within the macropore space of BCP/BMP-2 constructs. The human ECFC-derived vessels anastomosed with the host vasculature and perfused vessels were visible in the very center of the 5mm diameter, 2.5mm tall scaffolds. Additionally, the vessels were evenly distributed throughout the construct. This study suggests that scaffolds containing ECFCs have significant potential for expedited neovascularization in bony defects.
Acta biomaterialia 07/2011; 7(12):4222-8. · 3.98 Impact Factor
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ABSTRACT: Osteoinductive agents, such as BMP-2, are known to improve bone formation when combined with scaffolds. Microporosity (<20 μm) has also been shown to influence bone regeneration in calcium phosphate (CaP) scaffolds. However, many studies use only the term "osteoconductive" to describe the effects of BMP-2 and microporosity on bone formation, and do not assess the degree of healing that occurred. The objective of this study was to quantify the influence of BMP-2 and microporosity on bone regeneration and healing in biphasic calcium phosphate scaffolds using multiple measures including bone volume fraction, radial distribution, and specific surface area. These measures were quantitatively compared by analyzing microcomputed tomography data and used to formally define and assess healing. A custom image segmentation program was used to segment >100 samples, with 900 images each, that were implanted in porcine mandibular defects for 3, 6, 12 and 24 weeks. The assessment of healing presented in this work demonstrates the level of detail possible in evaluating scaffold-guided bone regeneration. The analysis shows that BMP-2 and microporosity accelerate healing up to 4-fold. BMP-2 and microporosity were shown to have different and complementary roles in bone formation that effect the time needed for a defect to heal.
Acta biomaterialia 12/2010; 7(4):1760-71. · 3.98 Impact Factor
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ABSTRACT: The role of macropore size (>100 microm) and geometry in synthetic scaffolds for bone regeneration has been studied extensively, but successful translation to the clinic has been slow. Significantly less attention has been given to porosity at the microscale (0.5-10 microm). While some have shown that microporosity in calcium phosphate (CaP)-based scaffolds can improve rate and extent of bone formation in macropores, none has explored microporosity as an additional and important space for bone ingrowth. Here we show osteointegration of biphasic calcium phosphate (BCP) scaffolds at both the macro and micro length scales. Bone, osteoid, and osteogenic cells fill micropores in scaffold rods and osteocytes are embedded in mineralized matrix in micropores, without the addition of growth factors. This work further highlights the importance of considering design parameters at the microscale and demonstrates the possibility for a bone-scaffold composite with no "dead space." Embedded osteocytes distributed throughout microporous rods may form a mechanosensory network, which would not be possible in scaffolds without microporosity. Multiscale osteointegration has the potential to greatly improve overall performance of these scaffolds through an improvement of mechanical properties, load transfer, and stability in the long and short term, and represents a new paradigm for scaffold design.
Biomaterials 02/2010; 31(13):3552-63. · 7.40 Impact Factor
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ABSTRACT: It is well established that scaffolds for applications in bone tissue engineering require interconnected pores on the order of 100 microm for bone in growth and nutrient and waste transport. As a result, most studies have focused on scaffold macroporosity (>100 microm). More recently researchers have investigated the role of microporosity in calcium phosphate -based scaffolds. Osteointegration into macropores improves when scaffold rods or struts contain micropores, typically defined as pores less than approximately 50 microm. We recently demonstrated multiscale osteointegration, or growth into both macropores and intra-red micropores (<10 microm), of biphasic calcium phosphate (BCP) scaffolds. The combined effect of BMP-2, a potent osteoinductive growth factor, and multiscale porosity has yet to be investigated. In this study we implanted BCP scaffolds into porcine mandibular defects for 3, 6, 12 and 24 weeks and evaluated the effect of BMP-2 on multiscale osteointegration. The results showed that given this in vivo model BMP-2 influences osteointegration at the microscale, but not at the macroscale, but not at the macroscale. Cell density was higher in the rod micropores for scaffolds containing BMP-2 compared with controls at all time points, but BMP-2 was not required for bone formation in micropores. In contrast, there was essentially no difference in the fraction of bone in macropores for scaffolds with BMP-2 compared with controls. Additionally, bone in macropores seemed to have reached steady-state by 3 weeks. Multiscale osteointegration results in bone-scaffold composites that are fully osteointegrated, with no 'dead space'. These composites are likely to contain a continuous cell network as well as the potential for enhanced load transfer and improved mechanical properties.
Acta biomaterialia 02/2010; 6(8):3283-91. · 3.98 Impact Factor
