Laura A. Smith

Biomimetic nanofibrous gelatin/apatite composite scaffolds for bone tissue engineering

Mimicking certain features (e.g. nanoscale topography and biological cues) of natural extracellular matrix (ECM) is advantageous for the successful regeneration of damaged tissue. In this study, nanofibrous gelatin/apatite (NF-gelatin/apatite) composite scaffolds have been fabricated to mimic both the physical architecture and chemical composition of natural bone ECM. A thermally induced phase separation (TIPS) technique was developed to prepare nanofibrous gelatin (NF-gelatin) matrix. The NF-gelatin matrix mimicked natural collagen fibers and had an average fiber diameter of about 150nm. By integrating the TIPS method with porogen leaching, three-dimensional NF-gelatin scaffolds with well-defined macropores were fabricated. In comparison to Gelfoam (a commercial gelatin foam) with similar pore size and porosity, the NF-gelatin scaffolds exhibited a much higher surface area and mechanical strength. The surface area and compressive modulus of NF-gelatin scaffolds were more than 700 times and 10 times higher than that of Gelfoam, respectively. The NF-gelatin scaffolds also showed excellent biocompatibility and mechanical stability. To further enhance pre-osteoblast cell differentiation as well as improving mechanical strength, bone-like apatite particles (<2microm) were incorporated onto the surface of NF-gelatin scaffolds via a simulated body fluid (SBF) incubation process. The NF-gelatin/apatite scaffolds 5 days after SBF treatment showed significantly higher mechanical strength than NF-gelatin scaffolds 5 days after SBF treatment. Furthermore, the incorporated apatite in the NF-gelatin/apatite composite scaffold enhanced the osteogenic differentiation. The expression of BSP and OCN in the osteoblast-(NF-gelatin/apatite composite) constructs was about 5 times and 2 times higher than in the osteoblast-(NF-gelatin) constructs 4 weeks after cell culture. The biomimetic NF-gelatin/apatite scaffolds are, therefore, excellent for bone tissue engineering.

Nanostructured polymer scaffolds for tissue engineering and regenerative medicine

The structural features of tissue engineering scaffolds affect cell response and must be engineered to support cell adhesion, proliferation and differentiation. The scaffold acts as an interim synthetic extracellular matrix (ECM) that cells interact with prior to forming a new tissue. In this review, bone tissue engineering is used as the primary example for the sake of brevity. We focus on nanofibrous scaffolds and the incorporation of other components including other nanofeatures into the scaffold structure. Since the ECM is comprised in large part of collagen fibers, between 50 and 500 nm in diameter, well-designed nanofibrous scaffolds mimic this structure. Our group has developed a novel thermally induced phase separation (TIPS) process in which a solution of biodegradable polymer is cast into a porous scaffold, resulting in a nanofibrous pore-wall structure. These nanoscale fibers have a diameter (50-500 nm) comparable to those collagen fibers found in the ECM. This process can then be combined with a porogen leaching technique, also developed by our group, to engineer an interconnected pore structure that promotes cell migration and tissue ingrowth in three dimensions. To improve upon efforts to incorporate a ceramic component into polymer scaffolds by mixing, our group has also developed a technique where apatite crystals are grown onto biodegradable polymer scaffolds by soaking them in simulated body fluid (SBF). By changing the polymer used, the concentration of ions in the SBF and by varying the treatment time, the size and distribution of these crystals are varied. Work is currently being done to improve the distribution of these crystals throughout three-dimensional scaffolds and to create nanoscale apatite deposits that better mimic those found in the ECM. In both nanofibrous and composite scaffolds, cell adhesion, proliferation and differentiation improved when compared to control scaffolds. Additionally, composite scaffolds showed a decrease in incidence of apoptosis when compared to polymer control in bone tissue engineering. Nanoparticles have been integrated into the nanostructured scaffolds to deliver biologically active molecules such as growth and differentiation factors to regulate cell behavior for optimal tissue regeneration.

Enhancing Osteogenic Differentiation of Mouse Embryonic Stem Cells by Nanofibers

Controlled differentiation of embryonic stem cells (ESC) is necessary to their use as a cell source for tissue engineering or regeneration. To date, most studies have concentrated on chemical cues to direct ESC differentiation. However, during normal embryonic development, multiple factors beyond chemical cues play a role, including the extracellular matrix (ECM) in bone development. In this study, we use nanofibrous (NF) matrices to mimic the morphology of the ECM to examine the contribution of the ECM morphology to the differentiation of mouse ESC. After 12 h of differentiation culture, mouse ESC form protrusions interacting with NF matrices, while they appear not to interact with flat films. Immunofluorescence staining after 26 days of differentiation culture indicates a greater degree of differentiation for mouse ESC on NF matrices compared to flat films. Polymerase chain reaction results, also, show greater degree of osteogenic differentiation on NF matrices compared to flat films when osteogenic supplements are added to the culture. Overall, these results demonstrate that NF morphology contributes to the controlled differentiation of mouse ESC.

The Enhancement of human embryonic stem cell osteogenic differentiation with nano-fibrous scaffolding

Human embryonic stem cells (hESC) hold great promise as a cell source for tissue engineering since they possess the ability to differentiate into any cell type within the body. However, much work must still be done to control the differentiation of the hESC to the desired lineage. In this study, we examined the effects of the nanofibrous (NF) architecture in both two-dimensional (2-D) poly(l-lactic acid) (PLLA) thin matrices and 3-D PLLA scaffolds in vitro to assess their affect on the osteogenic differentiation of hESC in vitro compared to more traditional solid films and solid-walled (SW) scaffolds. In 2-D culture, hESC on NF thin matrices were found to express collagen type 1, Runx2, and osteocalcin mRNA of higher levels than the hESC on the solid films after 1 week of culture and increased mineralization was observed on the NF matrices compared to the solid films after 3 weeks of culture. After 6 weeks of 3-D culture, the hESC on the NF scaffolds expressed significantly more osteocalcin mRNA compared to these on the SW scaffolds. The data indicates that the NF architecture enhances the osteogenic differentiation of the hESC compared to more traditional scaffolding architecture.

Nano-structured polymer scaffolds for tissue engineering and regenerative medicine

The structural features of tissue engineering scaffolds affect cell response and must be engineered to support cell adhesion, proliferation and differentiation. The scaffold acts as an interim synthetic extracellular matrix (ECM) that cells interact with prior to forming a new tissue. In this review, bone tissue engineering is used as the primary example for the sake of brevity. We focus on nanofibrous scaffolds and the incorporation of other components including other nanofeatures into the scaffold structure. Since the ECM is comprised in large part of collagen fibers, between 50 and 500 nm in diameter, well-designed nanofibrous scaffolds mimic this structure. Our group has developed a novel thermally induced phase separation (TIPS) process in which a solution of biodegradable polymer is cast into a porous scaffold, resulting in a nanofibrous pore-wall structure. These nanoscale fibers have a diameter (50-500 nm) comparable to those collagen fibers found in the ECM. This process can then be combined with a porogen leaching technique, also developed by our group, to engineer an interconnected pore structure that promotes cell migration and tissue ingrowth in three dimensions. To improve upon efforts to incorporate a ceramic component into polymer scaffolds by mixing, our group has also developed a technique where apatite crystals are grown onto biodegradable polymer scaffolds by soaking them in simulated body fluid (SBF). By changing the polymer used, the concentration of ions in the SBF and by varying the treatment time, the size and distribution of these crystals are varied. Work is currently being done to improve the distribution of these crystals throughout three-dimensional scaffolds and to create nanoscale apatite deposits that better mimic those found in the ECM. In both nanofibrous and composite scaffolds, cell adhesion, proliferation and differentiation improved when compared to control scaffolds. Additionally, composite scaffolds showed a decrease in incidence of apoptosis when compared to polymer control in bone tissue engineering. Nanoparticles have been integrated into the nanostructured scaffolds to deliver biologically active molecules such as growth and differentiation factors to regulate cell behavior for optimal tissue regeneration.

Validity and Reliability of the Vestibular/Ocular Motor Screening and Associations With Common Concussion Screening Tools:

Background: Sustaining a concussion commonly results in vestibular impairments that may be associated with balance deficits. To screen for vestibular impairments after a concussion, the Vestibular/Ocular Motor Screening (VOMS) tool was developed. The relationship between the VOMS and other concussion screening tools, such as the Balance Error Scoring System (BESS) and King-Devick (K-D), have not been explored. Hypotheses: (1) VOMS would provide reliable results and not provoke symptoms in healthy adolescents and (2) VOMS test items would measure related aspects of vestibular function that are not measured through the BESS or K-D. Study Design: Cross-sectional, descriptive. Level of Evidence: Level 4. Methods: A total of 105 healthy adolescents (53 male, 52 female; mean age, 15.4 years) completed the VOMS, BESS, and K-D tests. A subsample of 21 adolescents (16 male, 5 female; mean age, 15.5 years) completed the VOMS twice. Results: The median total symptom score for all 7 VOMS items was 0 (0-5). The majority of the individual VOMS test items total symptom scores demonstrated a significant correlation with each other (rs = 0.25-0.66, P < 0.02). The individual VOMS items did not demonstrate a significant relationship to the BESS or K-D. VOMS items demonstrated high agreement in total symptom scores between testing trials, with near point convergence (NPC) distance demonstrating an intraclass correlation coefficient (ICC) of 0.95 (95% CI, 0.89-0.98; P < 0.001). The MDC95 (minimal detectable change with 95 confidence) for NPC distance was 4 cm. Conclusion: The VOMS did not provoke vestibular symptoms in healthy adolescents. The VOMS items measured unique aspects of vestibular function other than those measured by the BESS or K-D with good reliability. Clinical Relevance: Clinicians should consider implementing the VOMS as part of a comprehensive concussion assessment if vestibular impairment is suspected. If NPC distance is measured twice, a difference of >4 cm would be considered real change outside of measurement error.

Computer-designed nano-fibrous scaffolds

Nano-fibrous scaffolding mimics aspects of the extracellular matrix to improve cell function and tissue formation. Although several methods exist to fabricate nano-fibrous scaffolds, the combination of phase separation with reverse solid freeform fabrication (SFF) allows for scaffolds with features at three different orders of magnitude to be formed, which is not easily achieved with other nano-fiber fabrication methods. This technique allows for the external shape and internal pore structure to be precisely controlled in an easily repeatable manner, while the nano-fibrous wall architecture facilitates cellular attachment, proliferation, and differentiation of the cells. In this chapter, we examine the fabrication of computer-designed nano-fibrous scaffolds utilizing thermally induced phase separation and reverse SFF, and the benefits of such scaffolds over more traditional tissue engineering scaffolds on cellular function and tissue regeneration.

Reference values for the balance error scoring system in adolescents.

Abstract Objectives: Adolescents with mild traumatic brain injury (i.e. concussion) may experience postural stability impairments. The Balance Error Scoring System (BESS) is widely used in assessment of postural stability after concussion. Despite its common use in adolescents, the BESS lacks reference values in adolescents, limiting its clinical utility. The objective of this study is to report the reference values for the BESS in adolescents and to examine the effect of gender on the BESS scores. Methods: One hundred and ninety-one high school adolescents between the ages of 14–18 (M = 16.1, SD = 1.1) years of age completed the BESS. The effects of gender, age, body mass and height on the performance of BESS were examined. Additionally, the reported reference values for the BESS were stratified by gender. Results: Female participants demonstrated better performance on five of the six BESS conditions as well as the total error score (p < 0.001). No relationships were observed between age and body mass to the BESS scores. Conclusions: The effects of gender on the BESS performance support the gender-specific reference values reported in this study. These reference values provide benchmarks for clinicians when interpreting the BESS in the absence of individual baseline scores.

Nanostructured polymer scaffolds for tissue engineering and regenerative medicine: Nanostructured polymer scaffolds

The structural features of tissue engineering scaffolds affect cell response and must be engineered to support cell adhesion, proliferation and differentiation. The scaffold acts as an interim synthetic extracellular matrix (ECM) that cells interact with prior to forming a new tissue. In this review, bone tissue engineering is used as the primary example for the sake of brevity. We focus on nanofibrous scaffolds and the incorporation of other components including other nanofeatures into the scaffold structure. Since the ECM is comprised in large part of collagen fibers, between 50 and 500 nm in diameter, well-designed nanofibrous scaffolds mimic this structure. Our group has developed a novel thermally induced phase separation (TIPS) process in which a solution of biodegradable polymer is cast into a porous scaffold, resulting in a nanofibrous pore-wall structure. These nanoscale fibers have a diameter (50-500 nm) comparable to those collagen fibers found in the ECM. This process can then be combined with a porogen leaching technique, also developed by our group, to engineer an interconnected pore structure that promotes cell migration and tissue ingrowth in three dimensions. To improve upon efforts to incorporate a ceramic component into polymer scaffolds by mixing, our group has also developed a technique where apatite crystals are grown onto biodegradable polymer scaffolds by soaking them in simulated body fluid (SBF). By changing the polymer used, the concentration of ions in the SBF and by varying the treatment time, the size and distribution of these crystals are varied. Work is currently being done to improve the distribution of these crystals throughout three-dimensional scaffolds and to create nanoscale apatite deposits that better mimic those found in the ECM. In both nanofibrous and composite scaffolds, cell adhesion, proliferation and differentiation improved when compared to control scaffolds. Additionally, composite scaffolds showed a decrease in incidence of apoptosis when compared to polymer control in bone tissue engineering. Nanoparticles have been integrated into the nanostructured scaffolds to deliver biologically active molecules such as growth and differentiation factors to regulate cell behavior for optimal tissue regeneration.