Scapulohumeral kinematics and neuromuscular control during scaption are associated with passive stiffness and strength of periscapular muscles in competitive adolescent swimmers.

The passive stiffness and strength of periscapular muscles may affect scapulohumeral control, especially in overhead athletes due to sports-specific training. This study tried to assess the relationship between the passive stiffness and strength of periscapular muscles, scapulohumeral kinematics and neuromuscular control during scaption in swimmers. Ten male adolescent competitive front-crawl swimmers were recruited. The passive stiffness and strength of periscapular muscles were measured in standard postures by a hand-held myotonometer and dynamometer, respectively. Surface electromyography and electromagnetic tracking systems were synchronized to record the muscle activities and scapulohumeral kinematics during scaption. Correlations between the passive stiffness or strength of periscapular muscles and the kinematics or muscle activity were examined by Spearman's rank correlation coefficient. The maximal strength of periscapular muscles correlated positively with the ranges of upward and external rotation of the scapula and negatively with muscle activity during scaption. Passive stiffness of periscapular muscles was associated with the downward rotation of the scapula and triggered an increase in muscle activity. Increased passive stiffness or decreased strength in the periscapular muscles may affect their role in controlling the scapular rotation and contribute to compensation from adjacent muscles. Our findings suggest that when attempting to evaluate scapular behavior, it may be beneficial to examine muscle strength and passive stiffness of periscapular muscles.

Intraoperative Administration of Adipose Stromal Vascular Fraction Does Not Improve Functional Outcomes in Young Patients with Anterior Cruciate Ligament Reconstruction.

Adipose stromal vascular fraction (SVF) has a versatile cellular system for biologically augmented therapies. However, there have been no clinical studies investigating the benefits of the augmentation of anterior cruciate ligament reconstruction (ACLR) with SVF. We conducted a retrospective study in assessing the effects of intraoperative SVF administration on the functional outcomes in young patients with ACLR. The enrolled patients were divided into the control group (ACLR only) and the SVF group (ACLR with SVF). The functional outcomes in both groups were assessed by the Lysholm knee scoring system, the Tegner activity scale, and the International Knee Documentation Committee (IKDC) subjective evaluation form, and compared at several time points during a 12-month follow-up. We found that the sex distribution and pre-surgery scores were similar in the two groups, whereas the mean age of the SVF group was higher than that of the control group (p = 0.046). The between-group analysis and generalized estimating equation model analysis revealed that, while patients in the SVF group significantly improved all their functional outcomes at 12 months after surgery, this improvement was not significantly different from the results of patients in the control group (Lysholm, p = 0.553; Tegner, p = 0.197; IKDC, p = 0.486). No side effects were observed in either group. We concluded that the intraoperative administration of SVF does not improve or accelerate functional recovery after ACLR in young patients.

Synthesis, Characterization, and Electrospinning of a Functionalizable, Polycaprolactone-Based Polyurethane for Soft Tissue Engineering.

We synthesized a biodegradable, elastomeric, and functionalizable polyurethane (PU) that can be electrospun for use as a scaffold in soft tissue engineering. The PU was synthesized from polycaprolactone diol, hexamethylene diisocyanate, and dimethylolpropionic acid (DMPA) chain extender using two-step polymerization and designated as PU-DMPA. A control PU using 1,4-butanediol (1,4-BDO) as a chain extender was synthesized similarly and designated as PU-BDO. The chemical structure of the two PUs was verified by FT-IR and 1H-NMR. The PU-DMPA had a lower molecular weight than the PU-BDO (~16,700 Da vs. ~78,600 Da). The melting enthalpy of the PU-DMPA was greater than that of the PU-BDO. Both the PUs exhibited elastomeric behaviors with a comparable elongation at break (λ=L/L0= 13.2). The PU-DMPA had a higher initial modulus (19.8 MPa vs. 8.7 MPa) and a lower linear modulus (0.7 MPa vs. 1.2 MPa) and ultimate strength (9.5 MPa vs. 13.8 MPa) than the PU-BDO. The PU-DMPA had better hydrophilicity than the PU-BDO. Both the PUs displayed no cytotoxicity, although the adhesion of human umbilical artery smooth muscle cells on the PU-DMPA surface was better. Bead free electrospun PU-DMPA membranes with a narrow fiber diameter distribution were successfully fabricated. As a demonstration of its functionalizability, gelatin was conjugated to the electrospun PU-DMPA membrane using carbodiimide chemistry. Moreover, hyaluronic acid was immobilized on the amino-functionalized PU-DMPA. In conclusion, the PU-DMPA has the potential to be used as a scaffold material for soft tissue engineering.

Removal of an abluminal lining improves decellularization of human umbilical arteries.

The decellularization of long segments of tubular tissues such as blood vessels may be improved by perfusing decellularization solution into their lumen. Particularly, transmural flow that may be introduced by the perfusion, if any, is beneficial to removing immunogenic cellular components in the vessel wall. When human umbilical arteries (HUAs) were perfused at a transmural pressure, however, very little transmural flow was observed. We hypothesized that a watertight lining at the abluminal surface of HUAs hampered the transmural flow and tested the hypothesis by subjecting the abluminal surface to enzyme digestion. Specifically, a highly viscous collagenase solution was applied onto the surface, thereby restricting the digestion to the surface. The localized digestion resulted in a water-permeable vessel without damaging the vessel wall. The presence of the abluminal lining and its successful removal were also supported by evidence from SEM, TEM, and mechanical testing. The collagenase-treated HUAs were decellularized with 1% sodium dodecyl sulfate (SDS) solution under either rotary agitation, simple perfusion, or pressurized perfusion. Regardless of decellularization conditions, the decellularization of HUAs was significantly enhanced after the abluminal lining removal. Particularly, complete removal of DNA was accomplished in 24 h by pressurized perfusion of the SDS solution. We conclude that the removal of the abluminal lining can improve the perfusion-assisted decellularization.

Combination of inductive effect of lipopolysaccharide and in situ mechanical conditioning for forming an autologous vascular graft in vivo.

Autologous vascular grafts have the advantages of better biocompatibility and prognosis. However, previous studies that implanted bare polymer tubes in animals to grow autologous tubular tissues were limited by their poor yield rates and stability. To enhance the yield rate of the tubular tissue, we employed a design with the addition of overlaid autologous whole blood scaffold containing lipopolysaccharides (LPS). Furthermore, we applied in vivo dynamic mechanical stimuli through cyclically inflatable silicone tube to improve the mechanical properties of the harvested tissues. The effectiveness of the modification was examined by implanting the tubes in the peritoneal cavity of rats. A group without mechanical stimuli served as the controls. After 24 days of culture including 16 days of cyclic mechanical stimuli, we harvested the tubular tissue forming on the silicone tube for analysis or further autologous interposition vascular grafting. In comparison with those without cyclic dynamic stimuli, tubular tissues with this treatment during in vivo culture had stronger mechanical properties, better smooth muscle differentiation, and more collagen and elastin expression by the end of incubation period in the peritoneal cavity. The grafts remained patent after 4 months of implantation and showed the presence of endothelial and smooth muscle cells. This model shows a new prospect for vascular tissue engineering.

Preparation of aligned poly(glycerol sebacate) fibrous membranes for anisotropic tissue engineering.

The use of fibrous scaffolds for tissue repair or regeneration is advantageous for its microstructure similar to that of the native ECM. Aligned fibrous scaffold, in particular, can be used to manipulate cell alignment and hence the microstructure of the resultant tissue. In our previous study, nanofibers consisting of solely poly(glycerol sebacate) (PGS) have been successfully fabricated using core-shell coaxial electrospinning followed by curing and subsequent shell removal. When we tried to fabricate aligned PGS fibrous membranes by collecting the electrospun fibers on a rapidly rotating drum, however, loss of fibrous structure was observed upon curing. This might be due to the broken fibers that were collected under tension; the core PGS prepolymer that melts at high temperature could leak from the broken ends during curing. In this study, attempts were made to reduce the possibility of the fiber breakage. At each stage of preparation, fiber morphology was examined by SEM and fiber compositions were verified by Fourier transform infrared spectroscopy and differential scanning calorimetry. Mechanical properties of the aligned PGS fibrous membrane were evaluated by uniaxial tensile testing both in parallel and perpendicular to the principal fiber direction. SEM images showed that fibrous morphology was better preserved upon the adjustment of the shell composition and the rotational speed of the collector drum. The final PGS fibers remained to be aligned although the alignment was less strong than that of as-spun core-shell fibers. The aligned PGS fibrous membrane exhibited anisotropic mechanical properties with Young's modulus in parallel and perpendicular to the principal fiber direction being 0.98 ±â€¯0.04 MPa and 0.52 ±â€¯0.02 MPa, respectively. The aligned PGS fibrous membrane was capable of guiding the orientation of cultured cells and therefore has the potential to be used to fabricate structurally anisotropic tissue-engineered constructs.

Fabrication of a mechanically anisotropic poly(glycerol sebacate) membrane for tissue engineering.

Poly(glycerol sebacate) (PGS) has been used successfully as a scaffolding material for soft tissue engineering. PGS scaffolds, however, are usually mechanically isotropic, which may restrict their use in tissue repairs as many soft tissues in the body have anisotropic mechanical behaviors. Although various methods have been used to fabricate anisotropic scaffolds, it remains challenging to make anisotropic scaffolds from thermoset PGS. Here a new, simple method to fabricate an anisotropic PGS membrane which can then be used to construct thicker three-dimensional anisotropic scaffolds was developed. First, an aligned sacrificial poly(vinyl alcohol) fibrous membrane was prepared by electrospinning. The fibrous membrane was then partially immersed in PGS prepolymer solution, resulting in a composite membrane upon drying. After curing, the sacrificial fibers within the membrane were removed by water, supposedly leaving aligned cylindrical pores in the membrane. Both SEM and AFM illustrated aligned grooves on the surface of the resultant PGS membrane, indicating the successful removal of sacrificial fibers. The PGS membrane was validated to be mechanically anisotropic using uniaxial tensile testing along and perpendicular to the predominant pore direction. The in vitro cytocompatibility of the PGS membrane was confirmed. As a demonstration of its potential application in vascular tissue engineering, a tubular scaffold was constructed by wrapping a stack of two axisymmetric pieces of the anisotropic PGS membranes on a mandrel. The compliance of the scaffold was found to depend on the pitch angle of its double helical structure, imitating the anisotropic mechanical behavior of the arterial media. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 760-770, 2018.

Fabrication of poly(glycerol sebacate) fibrous membranes by coaxial electrospinning: Influence of shell and core solutions.

Although poly(glycerol sebacate) (PGS) has enjoyed great success in soft tissue engineering, it remains challenging to fabricate PGS fibers. In this study, coaxial electrospinning, in which polylactide (PLA) was used to confine and draw PGS prepolymer, was used to fabricate PGS fibrous membranes. Specifically, effects of adding poly(ethylene oxide) (PEO), which was removed prior to curing, in the shell were investigated. Transmission and scanning electron microscopy were used to confirm core-shell structure and morphology of fibers, respectively. Both the removal of PEO or PLA in the shell and the efficacy of PGS curing were verified by Fourier transform infrared spectroscopy and differential scanning calorimetry. Mechanical properties of the membranes with different shell and core contents were examined. We found that the addition of PEO to the shell reduced Young׳s modulus of the resulting cured membrane and increased its elongation at break significantly, the latter indicating better PGS curing. Moreover, with the addition of PEO, increasing PGS prepolymer concentration further increased the elongation at break and appeared to enhance the structural integrity of fibers; PGS fibrous membranes (with no PLA shell) were thus successfully fabricated after the removal of PLA. The Young׳s modulus of the PGS fibrous membrane was ~0.47MPa, which is similar to that of PGS solid sheets and some soft tissues. Finally, the cytocompatibility of the electrospun membranes was validated by Alamar blue and LDH assays.

Rapid Fabrication of a Cell-Seeded Collagen Gel-Based Tubular Construct that Withstands Arterial Pressure : Rapid Fabrication of a Gel-Based Media Equivalent.

Based on plastically compressed cell-seeded collagen gels, we fabricated a small-diameter tubular construct that withstands arterial pressure without prolonged culture in vitro. Specifically, to mimic the microstructure of vascular media, the cell-seeded collagen gel was uniaxially stretched prior to plastic compression to align collagen fibers and hence cells in the gel. The resulting gel sheet was then wrapped around a custom-made multi-layered braided tube to form aligned tubular constructs whereas the gel sheet prepared similarly but without uniaxial stretching formed control constructs. With the braided tube, fluid in the gel construct was further removed by vacuum suction aiming to consolidate the concentric layers of the construct. The construct was finally treated with transglutaminase. Both SEM and histology confirmed the absence of gaps in the wall of the construct. Particularly, cells in the wall of the aligned tubular construct were circumferentially aligned. The enzyme-mediated crosslinking increased burst pressure of both the constructs significantly; the extent of the increase of burst pressure for the aligned tubular construct was greater than that for the control counterpart. Increasing crosslinking left the compliance of the aligned tubular construct unchanged but reduced that of the control construct. Cells remained viable in transglutaminase-treated plastically compressed gels after 6 days in culture. This study demonstrated that by combining stretch-induced fiber alignment, plastic compression, and enzyme-mediated crosslinking, a cell-seeded collagen gel-based tubular construct with potential to be used as vascular media can be made within 3 days.

On the decellularization of fresh or frozen human umbilical arteries: implications for small-diameter tissue engineered vascular grafts.

Most tissues, including those to be decellularized for tissue engineering applications, are frozen for long term preservation. Such conventional cryopreservation has been shown to alter the structure and mechanical properties of tissues. Little is known, however, how freezing affects decellularization of tissues. The purpose of this study was two-fold: to examine the effects of freezing on decellularization of human umbilical arteries (HUAs), which represent a potential scaffolding material for small-diameter tissue-engineered vascular grafts, and to examine how decellularization affects the mechanical properties of frozen HUAs. Among many decellularization methods, hypotonic sodium dodecyl sulfate solution was selected as the decellularizing agent and tested on fresh HUAs to optimize decellularization conditions. The efficiency of decellularization was evaluated by DNA assay and histology every 12 up to 48 h. The optimized decellularization protocol was then performed on frozen HUAs. The stiffness, burst pressure, and suture retention strength of fresh HUAs and frozen HUAs before and after decellularization were also examined. It appeared that freezing decreased the efficiency of decellularization, which may be attributed to the condensed extracellular matrix caused by freezing. While the stiffness of fresh HUAs did not change significantly after decellularization, decellularization reduced the compliance of frozen HUAs. Interestingly, the stiffness of decellularized frozen HUAs was similar to that of decellularized fresh HUAs. Although little difference in stiffness was observed, we suggest avoiding freezing if more efficient and complete decellularization is desired.