Roles of mucus adhesion and cohesion in cough clearance.

Clearance of intrapulmonary mucus by the high-velocity airflow generated by cough is the major rescue clearance mechanism in subjects with mucoobstructive diseases and failed cilial-dependent mucus clearance, e.g., subjects with cystic fibrosis (CF) or chronic obstructive pulmonary disease (COPD). Previous studies have investigated the mechanical forces generated at airway surfaces by cough but have not considered the effects of mucus biophysical properties on cough efficacy. Theoretically, mucus can be cleared by cough from the lung by an adhesive failure, i.e., breaking mucus-cell surface adhesive bonds and/or by cohesive failure, i.e., directly fracturing mucus. Utilizing peel-testing technologies, mucus-epithelial surface adhesive and mucus cohesive strengths were measured. Because both mucus concentration and pH have been reported to alter mucus biophysical properties in disease, the effects of mucus concentration and pH on adhesion and cohesion were compared. Both adhesive and cohesive strengths depended on mucus concentration, but neither on physiologically relevant changes in pH nor bicarbonate concentration. Mucus from bronchial epithelial cultures and patient sputum samples exhibited similar adhesive and cohesive properties. Notably, the magnitudes of both adhesive and cohesive strength exhibited similar velocity and concentration dependencies, suggesting that viscous dissipation of energy within mucus during cough determines the efficiency of cough clearance of diseased, hyperconcentrated, mucus. Calculations of airflow-induced shear forces on airway mucus related to mucus concentration predicted substantially reduced cough clearance in small versus large airways. Studies designed to improve cough clearance in subjects with mucoobstructive diseases identified reductions of mucus concentration and viscous dissipation as key therapeutic strategies.

Atomic Layer Deposition of Hafnium(IV) Oxide on Graphene Oxide: Probing Interfacial Chemistry and Nucleation by using X-ray Absorption and Photoelectron Spectroscopies.

Interfacing graphene with metal oxides is of considerable technological importance for modulating carrier density through electrostatic gating as well as for the design of earth-abundant electrocatalysts. Herein, we probe the early stages of the atomic layer deposition (ALD) of HfO2 on graphene oxide using a combination of C and O K-edge near-edge X-ray absorption fine structure spectroscopies and X-ray photoelectron spectroscopy. Dosing with water is observed to promote defunctionalization of graphene oxide as a result of the reaction between water and hydroxyl/epoxide species, which yields carbonyl groups that further react with migratory epoxide species to release CO2 . The carboxylates formed by the reaction of carbonyl and epoxide species facilitate binding of Hf precursors to graphene oxide surfaces. The ALD process is accompanied by recovery of the π-conjugated framework of graphene. The delineation of binding modes provides a means to rationally assemble 2D heterostructures.

Biochemical properties of tissue-engineered cartilage.

OBJECTIVE: Microtia is treated with rib cartilage sculpting and staged procedures; though aesthetically pleasing, these constructs lack native ear flexibility. Tissue-engineered (TE) elastic cartilage may bridge this gap; however, TE cartilage implants lead to hypertrophic changes with calcification and loss of flexibility. Retaining flexibility in TE cartilage must focus on increased elastin, maintained collagen II, decreased collagen X, with prevention of calcification. This study compares biochemical properties of human cartilage to TE cartilage from umbilical cord mesenchymal stem cells (UCMSCs). Our goal is to establish a baseline for clinically useful TE cartilage. METHODS: Discarded cartilage from conchal bowl, microtic ears, preauricular tags, rib, and TE cartilage were evaluated for collagen I, II, X, calcium, glycosaminoglycans, elastin, and fibrillin I and III. Human UCMSCs were chondroinduced on 2D surfaces and 3D D,L-lactide-co-glycolic acid (PLGA) fibers. RESULTS: Cartilage samples demonstrated similar staining for collagens I, II, and X, elastin, and fibrillin I and III, but differed from rib. TE pellets and PLGA-supported cartilage were similar to auricular samples in elastin and fibrillin I staining. TE samples were exclusively stained for fibrillin III. Only microtic samples demonstrated calcium staining. CONCLUSIONS: TE cartilage expressed similar levels of elastin, fibrillin I, and collagens I and X when compared to native cartilage. Microtic cartilage demonstrated elevated calcium, suggesting this abnormal tissue may not be a viable cell source for TE cartilage. TE cartilage appears to recapitulate the embryonic development of fibrillin III, which is not expressed in adult tissue, possibly providing a strategy to control TE elastic cartilage phenotype.

Physical Treatments and Therapies for Androgenetic Alopecia.

Androgenetic alopecia, the most common cause of hair loss affecting both men and women, is typically treated using pharmaceutical options, such as minoxidil and finasteride. While these medications work for many individuals, they are not suitable options for all. To date, the only non-pharmaceutical option that the United States Food and Drug Administration has cleared as a treatment for androgenetic alopecia is low-level laser therapy (LLLT). Numerous clinical trials utilizing LLLT devices of various types are available. However, a myriad of other physical treatments for this form of hair loss have been reported in the literature. This review evaluated the effectiveness of microneedling, pulsed electromagnetic field (PEMF) therapy, low-level laser therapy (LLLT), fractional laser therapy, and nonablative laser therapy for the treatment of androgenetic alopecia (AGA). It also explores the potential of multimodal treatments combining these physical therapies. The majority of evidence in the literature supports LLLT as a physical therapy for androgenetic alopecia. However, other physical treatments, such as nonablative laser treatments, and multimodal approaches, such as PEMF-LLLT, seem to have the potential to be equally or more promising and merit further exploration.

The Use of 2-Octylcyanoacrylate Dressing in Reducing the Risk of Superficial Surgical Site Infections in Colorectal Stoma Surgery: A Retrospective Cohort Study.

BACKGROUND: Superficial surgical site infection (SSI) is a common morbidity following bowel resection surgery involving stoma formation with clinical and financial implications. The study aimed to evaluate the role of topical skin adhesive, 2-octylcyanoacrylate (Dermabond®) (2-OCA) in reducing wound infections following colorectal stoma surgery. METHODS: We performed a retrospective, single-centre, cohort study using clinical notes. All patients, over the age of 18, undergoing bowel resection either elective or emergency, with stoma formation over five years from January 2015 to December 2019 were included. The primary endpoint was SSI, defined by the clinical manifestation of inflammation including pain, erythema, and discharge, regardless of the microbiological culture results. Patients received either 2-OCA glue as wound dressing or standard firm adhesive wound dressing e.g. Opsite. RESULTS:  Overall, 604 patients were included in the study. The median age was 67; 187 (31%) patients received Dermabond (Group 1) and 417 (69%) received standard care (Group 2). A total of 288 (47%) patients were female, 134 (22%) had body mass index (BMI) greater than 30, 87 (14%) were diabetic, and 90 (15%) were smokers. A total of 279 (46%) patients had an American Society of Anesthesiologists (ASA) score of 3 and 4; 282 (47%) patients went through emergency surgery, 279 (64%) patients underwent dirty surgery, and 220 (35%) patients developed SSI. BMI greater than 30 compared to < 30 (OR: 2.32, 95% CI: 1.54-3.49, p<0.0001), diabetes compared to no diabetes (OR: 0.54, 95% CI: 0.32-0.92, p<0.0241), dirty surgery compared to clean surgery (OR: 2.26, 95% CI: 1.51-3.37, p<0.0001) and standard care, no 2-OCA glue use compared to the use of 2-OCA glue (OR: 1.52, 95% CI: 1.03-2.24, p=0.0343) were associated with SSIs.  Conclusion: Our study demonstrates that there is an association between 2-OCA and reduced SSIs in bowel resection surgery involving stoma formation when compared to standard methods of wound dressing. Further randomised clinical trials are recommended to strengthen this evidence and demonstrate causation.

The caBIG® Life Science Business Architecture Model.

MOTIVATION: Business Architecture Models (BAMs) describe what a business does, who performs the activities, where and when activities are performed, how activities are accomplished and which data are present. The purpose of a BAM is to provide a common resource for understanding business functions and requirements and to guide software development. The cancer Biomedical Informatics Grid (caBIG®) Life Science BAM (LS BAM) provides a shared understanding of the vocabulary, goals and processes that are common in the business of LS research. RESULTS: LS BAM 1.1 includes 90 goals and 61 people and groups within Use Case and Activity Unified Modeling Language (UML) Diagrams. Here we report on the model's current release, LS BAM 1.1, its utility and usage, and plans for future use and continuing development for future releases. AVAILABILITY AND IMPLEMENTATION: The LS BAM is freely available as UML, PDF and HTML (https://wiki.nci.nih.gov/x/OFNyAQ).

Gauging treatment impact: The development of exposure variables in a large-scale evaluation study.

While guidance on how to design rigorous evaluation studies abounds, prescriptive guidance on how to include critical process and context measures through the construction of exposure variables is lacking. Capturing nuanced intervention dosage information within a large-scale evaluation is particularly complex. The Building Infrastructure Leading to Diversity (BUILD) initiative is part of the Diversity Program Consortium, which is funded by the National Institutes of Health. It is designed to increase participation in biomedical research careers among individuals from underrepresented groups. This chapter articulates methods employed in defining BUILD student and faculty interventions, tracking nuanced participation in multiple programs and activities, and computing the intensity of exposure. Defining standardized exposure variables (beyond simple treatment group membership) is crucial for equity-focused impact evaluation. Both the process and resulting nuanced dosage variables can inform the design and implementation of large-scale, diversity training program, outcome-focused, evaluation studies.

Single scan PC-MRI by alternating the velocity encoding gradient polarity between phase encoding steps.

The purpose of this study was to develop a faster approach to phase contrast magnetic resonance imaging. This article proposes a phase contrast imaging scheme called single scan phase contrast in which the polarity of the velocity-encoding gradient is alternated between phase encoding steps. In single scan phase contrast, ghost images due to moving spins form. The signal intensity of the ghost images is modulated by the sine of the motion-induce phase shift. Prior to image acquisition, the region of interest containing moving spins is identified, and the field of view is configured so to avoid overlap between the object in the image and the ghost image(s) due to motion in the region of interest. The image values of the region of interest and the ghost image are used to quantify velocity. At best, single scan phase contrast reduces the total acquisition time by a factor of two when compared to phase contrast. In this study, single scan phase contrast is validated against phase contrast in phantom and in vivo.

Bioengineered internal anal sphincter derived from isolated human internal anal sphincter smooth muscle cells.

BACKGROUND & AIMS: The internal anal sphincter (IAS) is a specialized circular smooth muscle that maintains rectoanal continence. In vitro models are needed to study the pathophysiology of human IAS disorders. We bioengineered sphincteric rings from human IAS smooth muscle cells (SMC) and investigated their response to cholinergic stimulation as well as investigated whether protein kinase C (PKC) and Rho kinase signaling pathways remain functional. METHODS: 3-Dimensional bioengineered ring (3DBR) model of the human IAS was constructed from isolated human IAS SMC obtained from surgery. Contractile properties and force generation in response to acetylcholine, PKC inhibitor calphostin-C, Rho/ROCK inhibitor Y-27632, permeable Rho/ROCK inhibitor c3-exoenzyme, and PKC activator PdBU was measured. RESULTS: The human IAS 3DBR has the essential characteristics of physiologically functional IAS; it generated a spontaneous myogenic basal tone, and the constructs were able to relax in response to relaxants and contract in response to contractile agents. The constructs generated dose-dependent force in response to acetylcholine. Basal tone was significantly reduced by calphostin-C but not with Y-27632. Acetylcholine-induced force generation was also significantly reduced by calphostin-C but not with Y-27632. PdBU generated force that was equal in magnitude to acetylcholine. Thus, calphostin-C inhibited PdBU-induced force generation, whereas Y-27632 and c3 exoenzyme did not. CONCLUSIONS: These data indicate that basal tone and acetylcholine-induced force generation depend on signaling through the PKC pathway in human IAS; PKC-mediated force generation is independent of the Rho/ROCK pathway. This human IAS 3DBR model can be used to study the pathophysiology associated with IAS malfunctions.

Acceptability of UC781 gel as a rectal microbicide among HIV-uninfected women and men.

We studied the overall acceptability of UC781 gel formulation when applied rectally. Ten women and twenty-six men, all HIV-uninfected, were enrolled in a Phase 1, randomized, blinded, placebo-controlled safety and acceptability study of the vaginal microbicide gel UC781 applied rectally. Participants were randomized to three groups: 0.1% UC781 gel, 0.25% UC781 gel, or a placebo gel. Acceptability was assessed using structured questionnaires and qualitative in-depth interviews. After using UC781 gel rectally for seven consecutive days, participants' reports suggest that a UC781 gel formulation is highly acceptable and comparable to a placebo gel. The gels received favorable ratings overall and on attributes such as color, smell and consistency. All of the participants reported high intentions to use a gel like the one they used in this study. Acceptability research is essential in early phases of microbicide development to identify potential problems, understand user preferences, and introduce changes if needed.

Methodological Problems With Online Concussion Testing.

Reaction time testing is widely used in online computerized concussion assessments, and most concussion studies utilizing the metric have demonstrated varying degrees of difference between concussed and non-concussed individuals. The problem with most of these online concussion assessments is that they predominantly rely on consumer grade technology. Typical administration of these reaction time tests involves presenting a visual stimulus on a computer monitor and prompting the test subject to respond as quickly as possible via keypad or computer mouse. However, inherent delays and variabilities are introduced to the reaction time measure by both computer and associated operating systems that the concussion assessment tool is installed on. The authors hypothesized systems that are typically used to collect concussion reaction time data would demonstrate significant errors in reaction time measurements. To remove human bias, a series of experiments was conducted robotically to assess timing errors introduced by reaction time tests under four different conditions. In the first condition, a visual reaction time test was conducted by flashing a visual stimulus on a computer monitor. Detection was via photodiode and mechanical response was delivered via computer mouse. The second condition employed a mobile device for the visual stimulus, and the mechanical response was delivered to the mobile device's touchscreen. The third condition simulated a tactile reaction time test, and mechanical response was delivered via computer mouse. The fourth condition also simulated a tactile reaction time test, but response was delivered to a dedicated device designed to store the interval between stimulus delivery and response, thus bypassing any problems hypothesized to be introduced by computer and/or computer software. There were significant differences in the range of responses recorded from the four different conditions with the reaction time collected from visual stimulus on a mobile device being the worst and the device with dedicated hardware designed for the task being the best. The results suggest that some of the commonly used visual tasks on consumer grade computers could be (and have been) introducing significant errors for reaction time testing and that dedicated hardware designed for the reaction time task is needed to minimize testing errors.

Adhesive and Cohesive Peel Force Measurement of Human Airway Mucus.

In health, the high-speed airflow associated with cough represents a vital backup mechanism for clearing accumulated mucus from our airways. However, alterations in the mucus layer in cystic fibrosis (CF) and chronic obstructive pulmonary disease (COPD) leads to the mucus layer adhered to the airway surfaces, representing the nidus of chronic lung infection. To understand what is different about diseased mucus and why cough clearance is defective, there is a need for techniques to quantify the strength of the interactions limiting the ability of airflow to strip mucus from the airway surface (i.e., adhesive strength) or tear mucus apart (i.e., cohesive strength). To overcome the issues with measuring these properties in a soft (i.e., low elastic modulus) mucus layer, we present here novel peel-testing technologies capable of quantifying the mucus adhesive strength on cultured airway cells and cohesive strength of excised mucus samples. While this protocol focuses on measurements of airway mucus, this approach can easily be adapted to measuring adhesive/cohesive properties of other soft biological materials.

A quantitative method for determining spatial discriminative capacity.

BACKGROUND: The traditional two-point discrimination (TPD) test, a widely used tactile spatial acuity measure, has been criticized as being imprecise because it is based on subjective criteria and involves a number of non-spatial cues. The results of a recent study showed that as two stimuli were delivered simultaneously, vibrotactile amplitude discrimination became worse when the two stimuli were positioned relatively close together and was significantly degraded when the probes were within a subject's two-point limen. The impairment of amplitude discrimination with decreasing inter-probe distance suggested that the metric of amplitude discrimination could possibly provide a means of objective and quantitative measurement of spatial discrimination capacity. METHODS: A two alternative forced-choice (2AFC) tracking procedure was used to assess a subject's ability to discriminate the amplitude difference between two stimuli positioned at near-adjacent skin sites. Two 25 Hz flutter stimuli, identical except for a constant difference in amplitude, were delivered simultaneously to the hand dorsum. The stimuli were initially spaced 30 mm apart, and the inter-stimulus distance was modified on a trial-by-trial basis based on the subject's performance of discriminating the stimulus with higher intensity. The experiment was repeated via sequential, rather than simultaneous, delivery of the same vibrotactile stimuli. RESULTS: Results obtained from this study showed that the performance of the amplitude discrimination task was significantly degraded when the stimuli were delivered simultaneously and were near a subject's two-point limen. In contrast, subjects were able to correctly discriminate between the amplitudes of the two stimuli when they were sequentially delivered at all inter-probe distances (including those within the two-point limen), and improved when an adapting stimulus was delivered prior to simultaneously delivered stimuli. CONCLUSION: Subjects' capacity to discriminate the amplitude difference between two vibrotactile stimulations was degraded as the inter-stimulus distance approached the limit of their two-point spatial discriminative capacity. This degradation of spatial discriminative capacity lessened when an adapting stimulus was used. Performance of the task, as well as improvement on the task with adaptation, would most likely be impaired if the cortical information processing capacity of a subject or subject population were systemically altered, and thus, the methods described could be effective measures for use in clinical or clinical research applications.

Proficiency in pole handling during Nordic walking influences exercise effectiveness in middle-aged and older adults.

Nordic walking (NW) is a total body version of walking increasingly used as a health-promoting activity by middle-aged and older adults. The present study examined the relationship between force exerted through the pole and physiological response during NW. In this non-randomized exercise trial, 17 participants comprising 8 middle-aged and older recreationally trained Nordic walkers (NWrec: 63.7 ± 8.1 years) and 9 experienced NW instructors (NWinstr: 57.5 ± 7.8 years) underwent outdoor ordinary walking (OW) and NW bouts as fast as possible for 12 minutes. Walking distance, speed, heart rate (HR), energy expenditure (METs and J/kg/m) and upper and lower limb muscle activities using surface electromyogram (EMG) were assessed. A pole with a built-in load cell measured force applied to the pole with peak pole force, pole contact time, % of pole contact time with respect to the gait cycle, and pole impulse derived. We conducted two-way analysis of covariance adjusted for age and BMI. There was a significant group and walking type interaction for walking distance and speed (P = 0.04), METs (P < 0.01), and HR (P = 0.04) with higher values in the NWinstr group during NW than OW. As expected, upper limb EMG activities increased (P < 0.01) with NW in both groups. All pole force measures were significantly higher in NWinstr than NWrec (P ≤ 0.01). Change in walking distance and speed were correlated with pole peak force (r = 0.67, P < 0.01) and pole impulse (r = 0.63, P = 0.01). Similarly, change in METs was associated with peak pole force (r = 0.66, P < 0.01) and pole impulse (r = 0.56, P = 0.02). These results indicate that planting the pole on the ground more forcefully and for longer periods to derive a driving force in NW enhances the effectiveness of the exercise and potentially the health-derived benefits.

Excitability of skeletal muscle during development, denervation, and tissue culture.

A quantitative understanding of the bulk excitability of skeletal muscle tissues is important for the design of muscle tissue bioreactor systems, implantable muscle stimulators, and other systems where electrical pulses are employed to elicit contractions in muscle tissue both in vitro and in vivo. The purpose of the present study is to systematically compare the excitability of mammalian (rat) skeletal muscle under a range of conditions (including neonatal development, denervation, and chronic in vivo stimulation of denervated muscle) and of self-organized muscle tissue constructs engineered in vitro from both primary cells and cell lines. Excitability is represented by rheobase (R(50), units = V/mm) and chronaxie (C(50), units = microseconds) values, with lower values for each indicating greater excitability. Adult skeletal muscle is the most excitable (R(50) ~ 0.29, C(50) ~ 100); chronically denervated whole muscles (R(50) ~ 2.54, C(50) ~ 690) and muscle engineered in vitro from cell lines (C2C12 + 10T1/2) (R(50) ~ 1.93, C(50) ~ 416) have exceptionally low excitability; muscle engineered in vitro from primary myocytes (R(50) ~ 0.99, C(50) ~ 496) has excitability similar to that of day 14 neonatal rat muscle (R(50) ~ 0.65, C(50) ~ 435); stimulated-denervated muscles retain excellent excitability when chronically electrically stimulated (R(50) ~ 0.40, C(50) ~ 100); and neonatal rat muscle excitability improves during the first 6 weeks of development, steadily approaching that of adult muscle.

Development of self-assembled, tissue-engineered ligament from bone marrow stromal cells.

The human anterior cruciate ligament is ruptured 200,000 times per year in the United States, resulting in medical costs of $1 billion. The standard treatment is patellar tendon autograft, but this treatment is suboptimal because of lengthy recovery time, arthritis, donor site morbidity, and degenerative joint disease. This study aimed to engineer scaffold-free ligament analogs from a clinically relevant cell source and to examine mechanical and histological properties of the resulting engineered tissue. Porcine bone marrow stromal cells were seeded on laminin-coated substrates with silk suture segments as anchor points. Cells developed into monolayers that subsequently delaminated and self-organized into cohesive rod-like tissues that were held in tension above the substrate. After 14 days of maturation, scanning electron microscopy revealed a well-organized extracellular matrix, aligned collagen fibers, and a collagen fibril diameter of 51.1+/-77 nm. Histological evaluation showed that constructs were composed of approximately 60% collagen. During tensile tests to failure, constructs had a stress of 2.11 +/- 0.13 MPa, a strain of 28.8 +/- 0.95%, a force of 0.26 +/- 0.02 N, and a tangent modulus of 15.4+/-1.04 MPa. Mechanically and histologically, engineered ligament resembled native embryonic connective tissue and had an ultimate stress approximately 15% of native adult mouse tissue.

Electrical stimulation prior to delayed reinnervation does not enhance recovery in muscles of rats.

PURPOSE: Prolonged denervation of skeletal muscles results in atrophy and poor recovery of motor function following delayed reinnervation. Electrical stimulation reduces denervation atrophy. We hypothesized that electrical stimulation of denervated extensor digitorum longus (EDL) muscles during a prolonged period between nerve axotomy and opportunity for reinnervation by motoneurons after nerve-repair would enhance the recovery of muscle mass, force and motor-function. METHODS: The EDL muscles of rats were denervated for 3.5 months by peroneal nerve axotomy, then repaired with an end-to-end neurorrhaphy, and allowed to recover for 6.5 months. During the period of denervation, some of the rats received a protocol of electrical stimulation that had previously been shown to dramatically attenuate the effects of denervation atrophy through 4 months. Other experimental groups included unoperated control muscles, denervated muscles, and axotomy followed immediately by nerve-repair. Final evaluations included walking track analysis, maximum force measured in situ by indirect stimulation of the nerve, and muscle mass. RESULTS: The hypothesis was not supported. Electrical stimulation during the period of denervation did not enhance recovery of muscle mass, force or motor function. CONCLUSION: The primary factors that inhibited reinnervation and recovery following delayed reinnervation were not alleviated by the electrical stimulation during the period of muscle denervation.

The implications of an incidental chronic lymphocytic leukaemia in a resection specimen for colorectal adenocarcinoma.

BACKGROUND: Colorectal cancer and B cell chronic lymphocytic leukaemia (CLL) have a significant incidence, which are increasing with the aging population. Evidence has been presented in the literature to suggest that the synchronous presentation of colorectal cancer and B cell CLL may be more than simply coincidental for these two common malignancies. We report an unusual case of a presumed B cell CLL diagnosed on the basis of histological analysis of lymph nodes recovered from a resection specimen for rectal adenocarcinoma. We considered aetiological factors which may have linked the synchronous diagnosis of the two malignancies and the potential implications for the natural history of the two malignancies on one another. CASE PRESENTATION: A 70-year-old male underwent low anterior resection with total mesorectal excision for a rectal adenocarcinoma. His co-morbid conditions were chronic obstructive airways disease and ischaemic heart disease. General examination revealed no lymphadenopathy. Full blood count, urea and electrolytes and liver function tests were all within normal limits. As well as confirming a pT3 N1 adenocarcinoma, histological analysis showed lymph nodes with an infiltrate of small lymphoid cells. Immunohistochemical studies showed these cells to be in keeping with B cell CLL. CONCLUSION: Whilst unable to identify any common aetiological factors in the two malignancies in our patient, immunosuppression and genetic abnormalities have been identified as possible bases for an observed epidemiological association between colorectal cancer and haematological malignancies. Examples such as our case of synchronous diagnosis of two malignancies in a patient are likely to increase with the aging population. The potential affects of one malignancy on the natural history of the other warrants further study. In our case, we considered that slow progression of the B cell CLL may increase the risk of recurrent rectal adenocarcinoma.

Biomimetic approach to cardiac tissue engineering: oxygen carriers and channeled scaffolds.

We report that the functional assembly of engineered cardiac muscle can be enhanced by oxygen supply provided by mechanisms resembling those in normal vascularized tissues. To mimic the capillary network, cardiomyocytes and fibroblasts isolated from the neonatal rat hearts were cultured on a highly porous elastomer with a parallel array of channels that were perfused with culture medium. To mimic oxygen supply by hemoglobin, culture medium was supplemented with a perfluorocarbon (PFC) emulsion; constructs perfused with unsupplemented culture medium served as controls. In PFC-supplemented medium, the decrease in the partial pressure of oxygen in the aqueous phase was only 50% of that in control medium (28 mmHg vs. 45 mmHg between the construct inlet and outlet at a flow rate of 0.1 mL/min). Consistently, constructs cultivated in the presence of PFC contained higher amounts of DNA and cardiac markers (troponin I, connexin-43) and had significantly better contractile properties as compared to control constructs. In both groups, electron microscopy revealed open channels and the presence of cells at the channel surfaces as well as within constructs. Improved properties of cardiac constructs could be correlated with the enhanced supply of oxygen to the cells, by a combined use of channeled scaffolds and PFC.

Electrical stimulation of denervated muscles of rats maintains mass and force, but not recovery following grafting.

PURPOSE: Denervated skeletal muscles lack contractile activity and subsequently lose mass and force generation. Prolonged periods of denervation prior to nerve-implant grafting limit the recovery of mass and force. We hypothesized that electrical stimulation during a period of denervation that maintains mass and force above the levels of denervated muscles enhances the recovery of mass and force following nerve-implant grafting. METHODS: The extensor digitorum longus (EDL) muscles of anesthetized rats were denervated, and a stimulator was implanted. Following 4 or 7 months of denervation, with or without electrical stimulation, the EDL muscles were removed, evaluated in vitro for mass and contractile properties, and then nerve-implant grafted into syngeneic rats. Unoperated, contralateral muscles were also evaluated and grafted. RESULTS: The hypothesis was not supported by the experimental data. Compared with values for 4- or 7-month denervated muscles, the stimulated-denervated muscles maintained higher mass and force, less prolonged time-to-peak tensions and half-relaxation times, and higher excitability. Nevertheless, the recovery of mass and force following grafting was not improved. CONCLUSION: The factors within long-term denervated muscles that hinder recovery following grafting appear to be related primarily to factors associated with the duration of denervation and not to the level of atrophy and weakness prior to grafting.