Impact of Passaging Primary Skeletal Muscle Cell Isolates on the Engineering of Skeletal Muscle.

Volumetric muscle loss (VML) is a clinical state that results in impaired skeletal muscle function. Engineered skeletal muscle can serve as a treatment for VML. Currently, large biopsies are required to achieve the cells necessary for the fabrication of engineered muscle, leading to donor-site morbidity. Amplification of cell numbers using cell passaging may increase the usefulness of a single muscle biopsy for engineering muscle tissue. In this study, we evaluated the impact of passaging cells obtained from donor muscle tissue by analyzing characteristics of in vitro cellular growth and tissue-engineered skeletal muscle unit (SMU) structure and function. Human skeletal muscle cell isolates from three separate donors (P0-Control) were compared with cells passaged once (P1), twice (P2), or three times (P3) by monitoring SMU force production and determining muscle content and structure using immunohistochemistry. Data indicated that passaging decreased the number of satellite cells and increased the population doubling time. P1 SMUs had slightly greater contractile force and P2 SMUs showed statistically significant greater force production compared with P0 SMUs with no change in SMU muscle content. In conclusion, human skeletal muscle cells can be passaged twice without negatively impacting SMU muscle content or contractile function, providing the opportunity to potentially create larger SMUs from smaller biopsies, thereby producing clinically relevant sized grafts to aid in VML repair.

Impact of Human Recombinant Irisin on Tissue-Engineered Skeletal Muscle Structure and Function.

Tissue engineering of exogenous skeletal muscle units (SMUs) through isolation of muscle satellite cells from muscle biopsies is a potential treatment method for acute volumetric muscle loss (VML). A current issue with this treatment process is the limited capacity for muscle stem cell (satellite cell) expansion in cell culture, resulting in a decreased ability to obtain enough cells to fabricate SMUs of appropriate size and structural quality and that produce native levels of contractile force. This study determined the impact of human recombinant irisin on the growth and development of three-dimensional (3D) engineered skeletal muscle. Muscle satellite cells were cultured without irisin (control) or with 50, 100, or 250 ng/mL of irisin supplementation. Light microscopy was used to analyze myotube formation with particular focus placed on the diameter and density of the monotubes during growth of the 3D SMU. Following the formation of 3D constructs, SMUs underwent measurement of maximum tetanic force to analyze contractile function, as well as immunohistochemical staining, to characterize muscle structure. The results indicate that irisin supplementation with 250 ng/mL significantly increased the average diameter of myotubes and increased the proliferation and differentiation of myoblasts in culture but did not have a consistent significant impact on force production. In conclusion, supplementation with 250 ng/mL of human recombinant irisin promotes the proliferation and differentiation of myotubes and has the potential for impacting contractile force production in scaffold-free tissue-engineered skeletal muscle.

Repairing Volumetric Muscle Loss with Commercially Available Hydrogels in an Ovine Model.

Volumetric muscle loss (VML) is the loss of skeletal muscle that exceeds the muscle's self-repair mechanism and leads to permanent functional deficits. In a previous study, we demonstrated the ability of our scaffold-free, multiphasic, tissue-engineered skeletal muscle units (SMUs) to restore muscle mass and force production. However, it was observed that the full recovery of muscle structure was inhibited due to increased fibrosis in the repair site. As such, novel biomaterials such as hydrogels (HGs) may have significant potential for decreasing the acute inflammation and subsequent fibrosis, as well as enhancing skeletal muscle regeneration following VML injury and repair. The goal of the current study was to assess the biocompatibility of commercially available poly(ethylene glycol), methacrylated gelatin, and hyaluronic acid (HA) HGs in combination with our SMUs to treat VML in a clinically relevant large animal model. An acute 30% VML injury created in the sheep peroneus tertius (PT) muscle was repaired with or without HGs and assessed for acute inflammation (incision swelling) and white blood cell counts in blood for 7 days. At the 7-day time point, HA was selected as the HG to use for the combined HG/SMU repair, as it exhibited a reduced inflammation response compared to the other HGs. Six weeks after implantation, all groups were assessed for gross and histological structural recovery. The results showed that the groups repaired with an SMU (SMU-Only and SMU+HA) restored muscle mass to greater degree than the groups with only HG and that the SMU groups had PT muscle masses that were statistically indistinguishable from its uninjured contralateral PT muscle. Furthermore, the HA HG, SMU-Only, and SMU+HA groups displayed notable efficacy in diminishing pro-inflammatory markers and showed an increased number of regenerating muscle fibers in the repair site. Taken together, the data demonstrates the efficacy of HA HG in decreasing acute inflammation and fibrotic response. The combination of HA and our SMUs also holds promise to decrease acute inflammation and fibrosis and increase muscle regeneration, advancing this combination therapy toward clinically relevant interventions for VML injuries in humans.

The potential of iRest in measuring the hand function performance of stroke patients.

BACKGROUND: Clinical scales such as Fugl-Meyer Assessment (FMA) and Motor Assessment Scale (MAS) are widely used to evaluate stroke patient's motor performance. However, there are several limitations with these assessment scales such as subjectivity, lack of repeatability, time-consuming and highly depend on the ability of the physiotherapy. In contrast, robot-based assessments are objective, repeatable, and could potentially reduce the assessment time. However, robot-based assessments are not as well established as conventional assessment scale and the correlation to conventional assessment scale is unclear. OBJECTIVE: This study was carried out to identify important parameters in designing tasks that efficiently assess hand function of stroke patients and to quantify potential benefits of robotic assessment modules to predict the conventional assessment score with iRest. METHODS: Twelve predictive variables were explored, relating to movement time, velocity, strategy, accuracy and smoothness from three robotic assessment modules which are Draw I, Draw Diamond and Draw Circle. Regression models using up to four predictors were developed to describe the MAS. RESULTS: Results show that the time given should be not too long and it would affect the trajectory error. Besides, result also shows that it is possible to use iRest in predicting MAS score. CONCLUSION: There is a potential of using iRest, a non-motorized device in predicting MAS score.

Portable and Reconfigurable Wrist Robot Improves Hand Function for Post-Stroke Subjects.

Rehabilitation robots have become increasingly popular for stroke rehabilitation. However, the high cost of robots hampers their implementation on a large scale. This paper implements the concept of a modular and reconfigurable robot, reducing its cost and size by adopting different therapeutic end effectors for different training movements using a single robot. The challenge is to increase the robot's portability and identify appropriate kinds of modular tools and configurations. Because literature on the effectiveness of this kind of rehabilitation robot is still scarce, this paper presents the design of a portable and reconfigurable rehabilitation robot and describes its use with a group of post-stroke patients for wrist and forearm training. Seven stroke subjects received training using a reconfigurable robot for 30 sessions, lasting 30 min per session. Post-training, statistical analysis showed significant improvement of 3.29 points (16.20%, p = 0.027) on the Fugl-Meyer assessment scale for forearm and wrist components. Significant improvement of active range of motion was detected in both pronation-supination (75.59%, p = 0.018) and wrist flexion-extension (56.12%, p = 0.018) after the training. These preliminary results demonstrate that the developed reconfigurable robot could improve subjects' wrist and forearm movement.

Micromanipulation accuracy in pointing and tracing investigated with a contact-free measurement system.

This study examines micromanipulation accuracy in pointing and in tracing a circle, using a novel contact-free measurement system. Three groups of subjects enable us to investigate the influence of age and micromanipulation expertise. The results show that, for all groups of subjects, a 10x magnification increases accuracy, but larger magnification does not improve it further. Expertise leads to reduced error, and grip force does not affect accuracy in the magnified condition.

Influence of visual feedback and speed on micromanipulation accuracy.

Accuracy in micromanipulation tasks is limited and it is important to identify various factors affecting it. This paper studies the effect of visual magnification, speed and handedness to micromanipulation accuracy using microscope and LCD screen for feedback. Magnification of visual feedback increases the accuracy, but large magnification does not provide further improvement beyond 16x. Further, we observed a trade off between speed and accuracy in tracing a circular path, i.e. faster speed reduces the speed control ability of the hand. Finally, dominant/non-dominant hand is found to affect accuracy in motion.

The role of posture, magnification, and grip force on microscopic accuracy.

While tremor has been studied extensively, the investigations thus far do not give detailed information on how the accuracy necessary for micromanipulations is affected while performing tasks in microsurgery and the life sciences. This paper systematically studies the effects of visual feedback, posture and grip force on the trial error and tremor intensity of subjects holding a forceps-like object to perform a pointing task. Results indicate that: (i) Arm support improves accuracy in tasks requiring fine manipulation and reduces tremor intensity in the 2-8 Hz region, but hand support does not provide the same effect; hence freedom of wrist movement can be retained without a significant increase in trial error. (ii) Magnification of up to x10 is critical to carry out accurate micromanipulations, but beyond that level, magnification is not the most important factor. (iii) While an appropriate grip force must be learned in order to grasp micro-objects, such as a needle, without damaging them, the level of grip force applied does not affect the endpoint accuracy.

Development of a microsurgery training system.

Surgeons require significant training to acquire sufficient dexterity and hand-eye coordination to manipulate objects skillfully under the microscope. This paper presents a computer-based real-time simulation of microsurgery as well as the hardware setup. It presents a realistic physics-based elastic suture and blood vessel model, fast collision detection techniques, suture insertion process and novel approach of a haptic forceps. The simulation environment demonstrates a complete vascular suturing system to train skills such as grasping, suture placement, needle insertion and knot-tying running at 500 Hz, sufficient for physical realism.