Prevention of recurrent radioulnar heterotopic ossification by combined indomethacin and a dermal/silicone sheet implant: case report.

A 27-year-old, right-handed man developed severe radioulnar synostosis at the distal radius 7 months after open reduction and internal fixation of his distal radius fracture. Heterotopic ossification formed at the radius fracture site, requiring excision of the heterotopic bone and plate removal. A bilayer, dermal substitute and silicone sheet was placed between the radius and ulna in the interosseous space to prevent recurrence of the heterotopic ossification. After surgery, the patient was treated with indomethacin 25 mg orally 3 times daily for 6 weeks. At 1 year after surgery, he has retained full pronation and near normal supination. Radiographs demonstrate no new heterotopic bone formation.

Force characteristics of in vivo tissue-engineered myocardial constructs using varying cell seeding densities.

Experiments have been successfully performed culminating in functional, vascularized, three-dimensional cardiac muscle tissue. Past experience in tissue engineering has led us to the understanding that cell seeding density plays a critical role in the formation and function of both in vitro and in vivo engineered tissues. Therefore, to improve upon the mechanics of this model and to facilitate the formation of myocardial tissue with improved functional performance, we sought to optimize the seeding density of cardiomyocytes in these constructs. Neonatal cardiac myocytes were isolated from 2-day-old Fischer 344 rat hearts. Silicone chambers containing fibrin gel were seeded with varying numbers of cardiac cells (1, 5, 10, and 20 million). Control chambers were prepared using fibrin gel alone. All of the chambers were then implanted around the femoral vessels of isogenic rats. Six constructs per cell seeding density group were implanted. Histological and immunohistochemical evaluation was performed via hematoxylin and eosin, von Gieson, and alpha-sarcomeric actin staining protocols. Linear contractile force measurements were obtained for each construct following 4 weeks of in vivo implantation. After an implantation period of 4 weeks, the newly formed cardiac constructs contained within the chambers were harvested. The femoral vessels within the constructs were found to be patent in all cases. With direct electrical stimulation, the constructs were able to generate an average active force that varied depending on their seeding density. Constructs with seeding densities of 1, 5, 10, and 20 million cells produced an average active force of 208, 241, 151, and 108 microN, respectively. The control constructs did not generate any active force on electrical stimulation. This study demonstrates the in vivo survival, vascularization, organization, and function of transplanted myocardial cells. It is also apparent that cell seeding density plays a direct role in the force generation and mechanical properties of these engineered constructs. Among different groups using varying cell seeding densities, we found that the group with 5 million cells generated maximum active force.

Neurotization improves contractile forces of tissue-engineered skeletal muscle.

Engineered functional skeletal muscle would be beneficial in reconstructive surgery. Our previous work successfully generated 3-dimensional vascularized skeletal muscle in vivo. Because neural signals direct muscle maturation, we hypothesized that neurotization of these constructs would increase their contractile force. Additionally, should neuromuscular junctions (NMJs) develop, indirect stimulation (via the nerve) would be possible, allowing for directed control. Rat myoblasts were cultured, suspended in fibrin gel, and implanted within silicone chambers around the femoral vessels and transected femoral nerve of syngeneic rats for 4 weeks. Neurotized constructs generated contractile forces 5 times as high as the non-neurotized controls. Indirect stimulation via the nerve elicited contractions of neurotized constructs. Curare administration ceased contraction in these constructs, providing physiologic evidence of NMJ formation. Histology demonstrated intact muscle fibers, and immunostaining positively identified NMJs. These results indicate that neurotization of engineered skeletal muscle significantly increases force generation and causes NMJs to develop, allowing indirect muscle stimulation.

Negating Tissue Contracture Improves Volume Maintenance and Longevity of In Vivo Engineered Tissues.

BACKGROUND: Engineering large, complex tissues in vivo requires robust vascularization to optimize survival, growth, and function. Previously, the authors used a "chamber" model that promotes intense angiogenesis in vivo as a platform for functional three-dimensional muscle and renal engineering. A silicone membrane used to define the structure and to contain the constructs is successful in the short term. However, over time, generated tissues contract and decrease in size in a manner similar to capsular contracture seen around many commonly used surgical implants. The authors hypothesized that modification of the chamber structure or internal surface would promote tissue adherence and maintain construct volume. METHODS: Three chamber configurations were tested against volume maintenance. Previously studied, smooth silicone surfaces were compared to chambers modified for improved tissue adherence, with multiple transmembrane perforations or lined with a commercially available textured surface. Tissues were allowed to mature long term in a rat model, before analysis. RESULTS: On explantation, average tissue masses were 49, 102, and 122 mg; average volumes were 74, 158 and 176 µl; and average cross-sectional areas were 1.6, 6.7, and 8.7 mm for the smooth, perforated, and textured groups, respectively. Both perforated and textured designs demonstrated significantly greater measures than the smooth-surfaced constructs in all respects. CONCLUSIONS: By modifying the design of chambers supporting vascularized, three-dimensional, in vivo tissue engineering constructs, generated tissue mass, volume, and area can be maintained over a long time course. Successful progress in the scale-up of construct size should follow, leading to improved potential for development of increasingly complex engineered tissues.

Implantation increases tensile strength and collagen content of self-assembled tendon constructs.

Tissue-engineered tendons, derived from an autologous cell source, have the potential to provide an ideal replacement graft that is biologically compatible and has the ability to adapt to the specific mechanical requirements of the in vivo environment. Scaffold-free tendon constructs have been successfully engineered in vitro. However, when compared against native tendons the constructs demonstrate both a lower tensile strength and collagen content. We hypothesized that the in vitro environment lacks certain environmental stimuli and that implantation in vivo would facilitate the maturation of engineered tissues. Using primary Achilles tendon fibroblasts from adult rats, self-organizing constructs were created in vitro. Tendon constructs were implanted subcutaneously into the groins of adult rats for 4 wk, while controls remained in vitro. Implanted constructs increased in stiffness by three orders of magnitude when compared with the in vitro controls (7,500 vs. 22.3 kPa). This increase in tangent modulus correlated with a significant increase in collagen content, as measured by hydroxyproline concentration, from 3.9% for the in vitro controls to 22.7% in the in vivo conditioned group. In addition, collagen fiber diameter increased from 22.0 to 75.4 nm as a result of in vivo implantation. The tensile strength and collagen content of in vivo conditioned constructs were similar to the values determined for neonatal rat tibialis anterior tendons.

Ipsilateral pedicle TRAM flaps for breast reconstruction: are they as safe as contralateral techniques?

INTRODUCTION: Many surgeons are using the ipsilateral rotation of the TRAM flap because of easier flap insetting and improved aesthetic outcomes; however studies evaluating these techniques are limited to case series without groups for comparison. Our purpose was to evaluate the safety of the ipsilateral technique through a retrospective cohort study. METHODS: We retrospectively evaluated consecutive contralateral and ipsilateral TRAM flaps for mastectomy breast reconstruction. The incidence of ischemia-related flap complications and the incidence of major and minor complications were compared between the two groups. Multiple logistic regression was used to assess the effect of rotation type on complication rates, while controlling for potential confounding variables. RESULTS: Comparing 58 ipsilateral to 32 contralateral pedicle TRAM patients, the rate of a major complication was 20.7% and 28.1%, respectively, p=0.425; a minor complication was 50.0% vs. 34.4%, respectively, p=0.153; and total ischemic-related flap complications were 22.4% and 25.0% respectively, p=0.781. When controlling for potential confounders, the type of pedicle TRAM flap did not have a significant effect on the above complications. Advanced patient age was significantly correlated with an increased incidence of major complications (OR=1.09, p=0.031) and ischemic flap complications (OR=1.11, p=0.016). CONCLUSION: The ipsilateral pedicle TRAM flap appears as safe as the contralateral technique for breast reconstruction. Given the increased pedicle length and preservation of both the medial inframammary fold and xiphoid subunit, we believe that the ipsilateral technique is preferred over the contralateral rotation.

Tissue engineering of an implantable bioartificial hemofilter.

The first step in the tissue engineering of an implantable bioartificial kidney is the development of an implant that produces ultrafiltrate to replace glomerular function. A fabricated device containing synthetic hollow hemofiltration fibers was placed around the femoral vascular pedicle in rats, which initiated new tissue formation with a mature and durable neocapillary bed. The transudate fluid produced by this newly formed capillary bed accumulated through the hollow fibers into a subcutaneous port to allow evaluation of the fluid. In its first phase, this study evaluated various hollow fibers and tissue induction processes by the measurement of fluid volume, urea nitrogen, and total protein continuously for 6 weeks. New tissues formed within the implants surrounding the fibers, and the vascular density, vessel sizes, and percent cross-sectional vascular area were assessed by means of histomorphometric analysis after 6 weeks. The volume of fluid formation correlated with both vascular density and fiber membrane surface area. The implant fluid-to-serum ratios demonstrated a permselective filtrate. In a second phase, platelet-derived growth factor and vascular endothelial growth factor versus carrier alone were infused directly into the implants for the first 4 weeks in vivo through osmotic pumps and followed up to 9 weeks. Cumulative implant fluid volumes were significantly greater in the growth factor-treated group than in control animals and were associated with greater numbers of small-caliber blood vessels. These results provide the initial proof of concept in developing a tissue-engineered hemofilter prototype on a small scale in a rodent model.