The Use of Collagen Methacrylate in Actuating Polyethylene Glycol Diacrylate-Acrylic Acid Scaffolds for Muscle Regeneration.

After muscle loss or injury, skeletal muscle tissue has the ability to regenerate and return its function. However, large volume defects in skeletal muscle tissue pose a challenge to regenerate due to the absence of regenerative elements such as biophysical and biochemical cues, making the development of new treatments necessary. One potential solution is to utilize electroactive polymers that can change size or shape in response to an external electric field. Poly(ethylene glycol) diacrylate (PEGDA) is one such polymer, which holds great potential as a scaffold for muscle tissue regeneration due to its mechanical properties. In addition, the versatile chemistry of this polymer allows for the conjugation of new functional groups to enhance its electroactive properties and biocompatibility. Herein, we have developed an electroactive copolymer of PEGDA and acrylic acid (AA) in combination with collagen methacrylate (CMA) to promote cell adhesion and proliferation. The electroactive properties of the CMA + PEGDA:AA constructs were investigated through actuation studies. Furthermore, the biological properties of the hydrogel were investigated in a 14-day in vitro study to evaluate myosin light chain (MLC) expression and metabolic activity of C2C12 mouse myoblast cells. The addition of CMA improved some aspects of material bioactivity, such as MLC expression in C2C12 mouse myoblast cells. However, the incorporation of CMA in the PEGDA:AA hydrogels reduced the sample movement when placed under an electric field, possibly due to steric hindrance from the CMA. Further research is needed to optimize the use of CMA in combination with PEGDA:AA as a potential scaffold for skeletal muscle tissue engineering.

In vitro studies of Annona muricata L. extract-loaded electrospun scaffolds for localized treatment of breast cancer.

This paper presents in vitro studies of the sustained release of Annona muricata leaf extracts (AME) from hybrid electrospun fibers for breast cancer treatment. Electrospun hybrid scaffolds were fabricated from crude AME extracts, poly(lactic-co-glycolic acid)/gelatin (PLGA/Ge) and pluronic F127. The physicochemical properties of the AME extract and scaffolds were studied. The antiproliferative effects of the scaffolds were also assessed on breast cancer (MCF-7 and MDA-MB-231) and non-tumorigenic breast (MCF10A) cell lines. Scanning electron microscope micrographs revealed a random network of micro- and submicron fibers. In vitro drug release profiles, governed by quasi-Fickian diffusion at pH 7.4 and non-Fickian super case II at pH 6.7, showed initial burst AME release from the PLGA/Ge-AME and PLGA/Ge-F127/AME fibers at pH 7.4, and burst release from PLGA/Ge-F127/AME (not observed from PLGA/Ge-AME) at pH 6.7. Then, a slower, sustained release of the remaining AME from the fibers, attributed to the onset of degradation of the PLGA/Ge backbone, was observed for the next 72 hr. The cumulative release of AME was 89.33 ± 0.73% (PLGA/Ge-AME) and 51.17 ± 7.96% (PLGA/Ge-F127/AME) at pH 7.4, and 9.27 ± 2.3% and 73.5 ± 4.5%, respectively, at pH 6.7. Pluronic F127 addition increased the drug loading capacity and prolonged the sustained AME release from the fibers. The released AME significantly inhibited the in vitro growth of the breast cancer cells more than the non-tumorigenic cells, due to the induction of apoptosis, providing evidence for using pluronic F127-containing electrospun fibers for sustained and localized AME delivery to breast cancer cells.

Characterization and optimization of a positively charged poly (ethylene glycol)diacrylate hydrogel as an actuating muscle tissue engineering scaffold.

Hydrogels have been used for many applications in tissue engineering and regenerative medicine due to their versatile material properties and similarities to the native extracellular matrix. Poly (ethylene glycol) diacrylate (PEGDA) is an ionic electroactive polymer (EAP), a material that responds to an electric field with a change in size or shape while in an ionic solution, that may be used in the development of hydrogels. In this study, we have investigated a positively charged EAP that can bend without the need of external ions. PEGDA was modified with the positively charged molecule 2-(methacryloyloxy)ethyl-trimethylammonium chloride (MAETAC) to provide its own positive ions. This hydrogel was then characterized and optimized for bending and cellular biocompatibility with C2C12 mouse myoblast cells. Studies show that the polymer responds to an electric field and supports C2C12 viability.

Optimizing C2C12 myoblast differentiation using polycaprolactone-polypyrrole copolymer scaffolds.

Advancements in tissue engineering and biomaterial development have the potential to provide a scalable solution to the problem of large-volume skeletal muscle defects. Previous research on the development of scaffolds for skeletal muscle regeneration has focused on strategies for increasing conductivity, which has improved satellite cell attachment and differentiation. However, these strategies usually increase scaffold stiffness, which some studies suggest may be detrimental to myoblast development. In this study, the polymers polypyrrole (PPy) and polycaprolactone (PCL) were synthesized together into a copolymer (PPy-PCL) designed to increase scaffold conductivity without significantly influencing stiffness. Different scaffold groups were fabricated via electrospinning, characterized, and assessed for their suitability for myoblast proliferation and differentiation. The groups included an aligned and random iteration of pure PCL, 10% PPy-PCL, 20% PPy-PCL, and 40% PPy-PCL. Only the 40% PPy-PCL group had a measureable conductivity, and the addition of PPy-PCL had no significant effect on the stiffness of the scaffolds. The PPy-PCL copolymer significantly increased the attachment of C2C12 myoblasts as compared to pure PCL scaffolds, but the concentration of PPy-PCL did not significantly alter cell attachment. In addition, scaffolds with PPy-PCL promoted myoblast differentiation to a greater extent than scaffolds made of PCL as measured by fusion index and number of nuclei per myotube. Aligned scaffolds were superior to random scaffolds in almost all measures. These results suggest that conductivity may not be the key factor in improving skeletal muscle scaffolds. Instead, cell attachment and aligned guidance cues may have a greater impact on myoblast differentiation. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 220-231, 2019.

Soft Robotic Manipulation and Locomotion with a 3D Printed Electroactive Hydrogel.

Electroactive hydrogels (EAH) that exhibit large deformation in response to an electric field have received great attention as a potential actuating material for soft robots and artificial muscle. However, their application has been limited due to the use of traditional two-dimensional (2D) fabrication methods. Here we present soft robotic manipulation and locomotion with 3D printed EAH microstructures. Through 3D design and precise dimensional control enabled by a digital light processing (DLP) based micro 3D printing technique, complex 3D actuations of EAH are achieved. We demonstrate soft robotic actuations including gripping and transporting an object and a bidirectional locomotion.

Characterization and optimization of actuating poly(ethylene glycol) diacrylate/acrylic acid hydrogels as artificial muscles.

Large volume deficiencies in skeletal muscle tissue fail to heal with conservative treatments, and improved treatment methods are needed. Tissue engineered scaffolds for skeletal muscle need to mimic the optimal environment for muscle development by providing the proper electric, mechanical, and chemical cues. Electroactive polymers, polymers that change in size or shape in response to an electric field, may be able to provide the optimal environment for muscle growth. In this study, an electroactive polymer made from poly(ethylene glycol) diacrylate (PEGDA) and acrylic acid (AA) is characterized and optimized for movement and biocompatibility. Hydrogel sample thickness, overall polymer concentration, and the ratio of PEGDA to AA were found to significantly impact the actuation response. C2C12 mouse myoblast cells attached and proliferated on hydrogel samples with various ratios of PEGDA to AA. Future experiments will produce hydrogel samples combined with aligned guidance cues in the form of electrospun fibers to provide a favorable environment for muscle development.

An overview of recent patents on musculoskeletal interface tissue engineering.

Interface tissue engineering involves the development of engineered grafts that promote integration between multiple tissue types. Musculoskeletal tissue interfaces are critical to the safe and efficient transmission of mechanical forces between multiple musculoskeletal tissues, e.g., between ligament and bone tissue. However, these interfaces often do not physiologically regenerate upon injury, resulting in impaired tissue function. Therefore, interface tissue engineering approaches are considered to be particularly relevant for the structural restoration of musculoskeletal tissues interfaces. In this article, we provide an overview of the various strategies used for engineering musculoskeletal tissue interfaces with a specific focus on the recent important patents that have been issued for inventions that were specifically designed for engineering musculoskeletal interfaces as well as those that show promise to be adapted for this purpose.

Poly(3,4-ethylenedioxythiophene) nanoparticle and poly(ɛ-caprolactone) electrospun scaffold characterization for skeletal muscle regeneration.

Injuries to peripheral nerves and/or skeletal muscle can cause scar tissue formation and loss of function. The focus of this article is the creation of a conductive, biocompatible scaffold with appropriate mechanical properties to regenerate skeletal muscle. Poly(3,4-ethylenedioxythiophene) (PEDOT) nanoparticles (Np) were electrospun with poly(ɛ-caprolactone) (PCL) to form conductive scaffolds. During electrospinning, ribboning, larger fiber diameters, and unaligned scaffolds were observed with increasing PEDOT amounts. To address this, PEDOT Np were sonicated prior to electrospinning, which resulted in decreased conductivity and increased mechanical properties. Multi-walled carbon nanotubes (MWCNT) were added to the 1:2 solution in an effort to increase conductivity. However, the addition of MWCNT had little effect on scaffold conductivity, and the elastic modulus and yield stress of the scaffold increased as a result. Rat muscle cells attached and were active on the 1-10, 1-2, 3-4, and 1-1 PCL-PEDOT scaffolds; however, the 3-4 scaffolds had the lowest level of metabolic activity. Although the scaffolds were cytocompatible, further development of the fabrication method is necessary to produce more highly aligned scaffolds capable of promoting skeletal muscle cell alignment and eventual regeneration.

Changes to the mechanical properties of the glenohumeral capsule during anterior dislocation.

The glenohumeral joint is the most frequently dislocated major joint in the body, and instability due to permanent deformation of the glenohumeral capsule is a common pathology. The corresponding change in mechanical properties may have implications for the ideal location and extent of plication, which is a common clinical procedure used to repair the capsule. Therefore, the objective of this study was to quantify the mechanical properties of four regions of the glenohumeral capsule after anterior dislocation and compare the properties to the normal glenohumeral capsule. Six fresh-frozen cadaveric shoulders were dislocated in the anterior direction with the joint in the apprehension position using a robotic testing system. After dislocation, mechanical testing was performed on the injured glenohumeral capsule by loading the tissue samples in tension and shear. An inverse finite element optimization routine was used to simulate the experiments and obtain material coefficients for each tissue sample. Cauchy stress-stretch curves were then generated to represent the mechanical response of each tissue sample to theoretical loading conditions. Based on several comparisons (average of the material coefficients, average stress-stretch curve for each region, and coefficients representing the average curves) between the normal and injured tissue samples, the mechanical properties of the injured tissue samples from multiple regions were found to be lower than those of the normal tissue in tension but not in shear. This finding indicates that anterior dislocation primarily affects the tensile behavior of the glenohumeral capsule rather than the shear behavior, and this phenomenon could be caused by plastic deformation of the matrix, permanent collagen fiber rotation, and/or collagen fiber failure. These results suggest that plication and suturing may not be sufficient to return stability to the shoulder after dislocation in all individuals. Thus, surgeons may need to perform a procedure that reinforces or stiffens the tissue itself, such as reconstruction or augmentation, to improve repair procedures.

Injury to the anteroinferior glenohumeral capsule during anterior dislocation.

BACKGROUND: Glenohumeral dislocation commonly results in permanent deformation of the glenohumeral capsule. Knowing the location and extent of tissue damage may aid in improving diagnostic and repair procedures for shoulder dislocations. Therefore, the objectives of this study were to determine: (1) the strain in the anteroinferior capsule at dislocation and (2) the location and extent of injury to the anteroinferior capsule due to dislocation by quantifying the resulting non-recoverable strain. METHODS: A robotic/universal force-moment sensor testing system was used to anteriorly dislocate six cadaveric shoulders. The magnitude of the maximum principle strain at dislocation and the resulting non-recoverable strain due to dislocation in the anteroinferior capsule were measured by tracking the change in the location of a grid of strain markers from a reference position. FINDINGS: The glenoid side of the capsule experienced higher strains at dislocation than the humeral side. The greatest strains at dislocation were found on the glenoid side of the anterior band (strain ratio of 0.60), but the greatest non-recoverable strains were found in the posterior axillary pouch (strain ratio of 0.34 on the glenoid side and 0.31 on the humeral side). INTERPRETATION: These findings suggest that even though the glenoid side of the anterior band undergoes more deformation during anterior dislocation, the most permanent deformation occurs in the posterior axillary pouch, and surgeons should consider also plicating the posterior axillary pouch when performing repair procedures following anterior dislocation. In the future, the mechanical properties of the normal and injured glenohumeral capsules will be compared.

Capsule function following anterior dislocation: implications for diagnosis of shoulder instability.

During shoulder dislocation, the glenohumeral capsule undergoes non-recoverable strain, leading to joint instability. Clinicians use physical exams to diagnose injury and direct repair procedures; however, they are subjective and do not provide quantitative information. Our objectives were to: (1) determine the relationship between capsule function following anterior dislocation and non-recoverable strain; and (2) identify joint positions at which physical exams can be used to detect non-recoverable strain in specific capsule regions. Physical exams were simulated at three joint positions including external rotation (ER) using robotic technology before and after anterior dislocation. The resulting joint kinematics, strain distribution in the capsule, and non-recoverable strain were determined. Following dislocation, anterior translation increased by as much as 48% (0° ER: p = 0.03; 30° ER: p = 0.03; 60° ER: p < 0.01). Capsule sub-regions with less non-recoverable strain required more ER to detect differences in the strain ratios between the intact and injured joint. Strain ratio changes on the humeral side of the posterior axillary pouch (0.31 ± 0.32) were significant at all joint positions (0° ER: p = 0.03; 30° ER: p = 0.048; 60° ER: p = 0.04), whereas strain ratio differences on the humeral side of the anterior axillary pouch (0.18 ± 0.21) were significant only at 60° of ER (p = 0.03). Therefore, standardizing physical exams for joint position could help surgeons identify specific locations of non-recoverable strain that may have been ignored.