Repeat sternotomy after reconstruction of the pericardial sac with glutaraldehyde-preserved equine pericardium

The risk of repeat sternotomy is higher than that of the initial sternotomy, especially if the pericardial sac was left open at the first intervention. In 200 consecutive patients with a pericardial defect after open heart operations, the pericardium was closed with a glutaraldehyde-preserved equine pericardial patch. Precardiac adhesions at reoperation were assessed in four groups of patients on a scale of 6, ranging from 0 (no adhesions) to 5 (calcified or ossified adhesions). Group I comprised 13 patients in whom the pericardium was left open at the first operation and an equine pericardial patch was implanted at reoperation. Group Ia included the first five Group I patients who underwent reoperation less than 1 year (early reoperation) after the initial procedure. Group Ib included the other eight patients of Group I, who underwent reoperation more than 1 year (late reoperation) after the first procedure. Group II comprised nine patients who were reoperated on after reconstruction of the pericardial sac with a glutaraldehyde-preserved equine pericardial patch. After a mean follow-up of 20.2 months, the incidence of patch-related complications was 1%. Statistical analysis shows less severe adhesions on reoperation in Group II patients (pericardial defect patched) than in Group I patients (pericardial defect left open): mean grade of adhesions 1.6 +/- 0.9 (Group II) versus 3.2 +/- 0.6 (Group I), p less than 0.001. Precardial adhesions with the pericardium left open were similar in patients having early and late reoperations: mean grade of adhesions 3.0 +/- 0.7 (Group Ia) versus 3.4 +/- 0.5 (Group Ib), no significant difference. Therefore, the glutaraldehyde-preserved equine pericardial patch can be considered a suitable material for primary closure of the pericardial sac in patients with inadequate autologous pericardium.     

Role of IL‐17 and TGF‐β in peritoneal adhesion formation after surgical trauma

Peritoneal adhesions are fibrous tissues formed after surgery. Both cytokines and transforming growth factors (TGFs) are involved in this process. The objective of this study was to investigate the cross talk between these entities. Peritoneal drainage fluid after surgery from patients and rodent models was examined by enzyme‐linked immunosorbent assay and fluorescence‐activated cell sorter. Data showed that the concentrations of interferon (IFN)‐γ and interleukin (IL)‐17 reached their peaks 6–12 hours after surgery, whereas TGF‐β1 concentrations showed two postoperative peak time points at 2 and 72–96 hours. By neutralizing IFN‐γ, IL‐17 6–12 hours, and TGF‐β1 72–96 hours after surgery, the degree of adhesion reduced significantly. However, neutralizing TGF‐β1 2 hours after surgery did not affect adhesion formation. Furthermore, in vitro studies showed that compared with the fibroblasts that were directly stimulated with TGF‐β1, the prestimulation of IL‐17 promoted plasminogen activator inhibitor‐1 production while inhibiting tissue‐type plasminogen activator production. Moreover, additional stimulation with IFN‐γ enhanced this effect. Together, these data indicate that IL‐17 may promote adhesion formation by increasing the reaction of fibroblasts against TGF‐β1. Blocking IL‐17 might have a therapeutic potential in preventing adhesion formation after surgery.     

Urinary transforming growth factor-β1 in children with obstructive uropathy

Aim: Obstructive uropathies (OU) in childhood constitute one of the major causes of chronic renal insufficiency. Transforming growth factor‐β1 (TGF‐β1) is considered to be the major fibrogenic growth factor. The aim of the present study was to investigate urinary TGF‐β1 levels in children with obstructive and non‐obstructive uropathies (NOU). Methods: This study involved 19 children with OU, 11 children with non‐obstructive hydronephrosis and 21 healthy children. Urinary TGF‐β1, proteinuria, microalbuminuria and urinary α1‐microglobulin were measured, and renal function was assesed. The results were statistically analyzed. Results: Mean urinary TGF‐β1 concentrations in patients with OU were significantly higher than those with NOU (4.14 ± 0.67 creatinine vs 1.80 ± 0.24 pg/mmol creatinine, P < 0.05) and healthy controls (1.66 ± 0.28 pg/mmol creatinine, P < 0.05). Positive correlations of urinary TGF‐β1 concentrations with proteinuria (r = 0.87, P < 0.0001) and urinary α1‐microglobulin (r = 0.82, P = 0.0002) were found in patients with OU. Conclusion: Children with OU have higher urinary TGF‐β1 than children with NOU. Urinary TGF‐β1 may be a useful non‐invasive tool for the differential diagnosis between OU and NOU in children. A positive correlation of TGF‐β1 with markers of renal tissue damage in patients with OU was found.     

Pericardial meshing: an effective method for prevention of pericardial adhesions and epicardial reaction after cardiac operations

Cardiac reoperations, particularly for coronary revascularization, are becoming more frequent and carry increased risk of damage to the heart during resternotomy. We experimentally evaluated a pericardial meshing technique to facilitate primary pericardial closure. In 18 mongrel dogs, an 8 by 5 cm pericardial flap was fashioned through a left thoracotomy. A standardized procedure for induction of pericardial adhesions was carried out in all animals. Animals were divided into three groups of six animals each: Group I (control)--the pericardial flap was primarily resutured; Group II--the flap was meshed and then resutured; and Group III--the flap was replaced by a pericardial substitute. Animals were put to death 8 weeks postoperatively and the pericardial space was examined for adhesions and epicardial reaction. The extent of adhesions and epicardial reaction was graded as: 0--none; 1--minimal; 2--moderate; and 3--severe. Both in Group I and Group III severe pericardial adhesions (grade 2-3) and epicardial reactions (grade 2-3) were formed, which obscured the underlying coronary anatomy. In Group II pericardial adhesions and epicardial reactions were none to minimal (grade 0-1) and the underlying coronary anatomy was not obscured. The meshed pericardium was completely regenerated by normal pericardium within several weeks. This study demonstrates that pericardial meshing facilitates primary tension-free pericardial closure. Free drainage of intrapericardial blood is achieved. A complete anatomic layer between heart and sternum is restored. Pericardial meshing is superior to the pericardial substitutes examined, as adhesions and epicardial reactions are significantly reduced, and the coronary anatomy is readily identifiable.     

A New Approach for Decreasing Postoperative Peritoneal Adhesions: Preventing Peritoneal Trauma with Soybean Oil

Background: Covering peritoneal surfaces with soybean oil may decrease peritoneal adhesions by preventing peritoneal trauma. Method(s): Forty female albino Wistar rats were divided into four equal groups. In Group 1, soybean oil only (0.1 ml) was injected into the peritoneal cavity. In Group 2, an untreated adhesion model was generated. In Group 3, an adhesion model was generated, followed by covering the area with soybean oil (0.1 ml). In Group 4, the area was first covered with soybean oil (0.1 ml) followed by generation of an adhesion model. All rats were sacrificed on postoperative day 10, and adhesions were scored. Results: The mean macroscopic adhesion scores in Groups 1, 2, 3, and 4 were 0.0 ± 0.0, 2.90 ± 0.21, 1.90 ± 0.94, and 0.50 ± 0.71, respectively. The Group 4 score differed significantly from that of Group 2 (p <. 001), but was not different from that of Group 1 or 3 (p >. 05). Discussion: Soybean oil can effectively decrease adhesion formation if applied before peritoneal trauma.     

Evaluation of a bioabsorable polylactide film in a large animal model for the reduction of retrosternal adhesions

Objectives

An adult pig model of retrosternal adhesion formation via an inferior hemisternotomy was used to evaluate the formation and development of pericardial and retrosternal adhesions, as well as adhesion reduction using two thicknesses of a bioabsorbable polylactide film.

Materials and methods

Twenty-five adult female pigs (70 kg) were allocated to either a control group or four different treatments using two thicknesses (0.02 or 0.05 mm) of a polylactide film. In each animal, the film was placed either inside the pericardium or inside and outside the pericardium.

Results

All animals demonstrated adhesions between the posterior and lateral surfaces of the heart and pericardium. Thick fibrous retrosternal adhesions and pericardial adhesions were noted in the control animals with complete obliteration of the anatomical plane. The polylactide films preserved the anatomical planes and reduced the adhesion response.

Conclusions

A reproducible animal model was used to examine the formation and reduction of retrosternal and pericardial adhesions. A polylactide film placed inside the pericardium or between the heart and sternum was able to limit adhesion formation and maintain the anatomical planes, which would facilitate reentry.     

Obstruction of St. Jude Medical Valves in the Aortic Position: Plasma Transforming Growth Factor Type Beta 1 in Patients With Pannus Overgrowth

The study investigated the hypothesis that plasma transforming growth factor type beta 1 (TGF‐β1) initiated pannus overgrowth in cases with aortic prosthetic valve dysfunction (PVD). Patients with obstruction of an aortic St. Jude Medical valve in 26 cases (PVD group) and without obstruction in 48 cases (control group) were studied. Plasma TGF‐β1, the intensity of the prothrombin time–international normalized ratio (PT‐INR), and the interruption of an oral anticoagulant medicine were conducted. Plasma TGF‐β1 levels in the PVD group (87.7 ± 29.2 ng/mL) were significantly higher (P < 0.05) than in the control group (73.7 ± 25.2 ng/mL). The interruption of an oral anticoagulant medicine in 54% of the PVD group versus 12% of the control group was identified (P < 0.001). The mean value of the PT‐INR in the PVD group (1.75 ± 0.30) and control group (1.75 ± 0.30) was not significantly different (P = 0.82). In conclusion, elevated levels of plasma TGF‐β1 may play a role in pannus overgrowth.     

Pathological Effects of Processed Bovine Pericardial Scaffolds—A Comparative In Vivo Evaluation

The objective of the present study was to assess the biocompatibility and regenerative potential of decellularized bovine pericardial scaffold in comparison with glutaraldehyde‐treated and fresh bovine pericardial implants using short‐term intramuscular implantation testing in a rat model. The inflammatory and immune responses were assessed using histopathological examination, special stains for connective tissue, histomorphometric evaluation, and immunohistochemistry. The decellularized pericardium showed an active tissue remodeling response with complete cellular invasion, minimum connective tissue encapsulation, extensive fibrovascular tissue formation, and collagen deposition. On the contrary, the glutaraldehyde‐treated pericardial implants showed incomplete degradation and cellular invasion, while the fresh pericardial implants elicited a severe foreign body reaction. The results of immunohistochemical staining revealed a minimum T helper (CD4+) lymphocyte response in decellularized pericardial implants compared with its glutaraldehyde‐treated and fresh counterparts. The decellularized bovine pericardium was better accepted as a prosthetic scaffold, which permitted maximum collagen deposition and active tissue remodeling by invading host cells and showed good tissue integration in vivo compared with glutaraldehyde‐treated and fresh/untreated pericardium.     

Is high levels of vitamin D a new risk factor for Peyronie's disease?

The aim of this study is to investigate whether serum vitamin D level predicts the risk of Peyronie's disease. Calcium and inflammatory cytokines play an important role during fibrocalcification of the plaques in Peyronie's Disease. TGF‐β1 is one of the most fibrogenic cytokines. Increasing serum vitamin D levels is considered that induce expression of TGF‐β1. Serum vitamin D levels and TGF‐β1 are related with calcifications of some soft tissues in previous studies. One hundred and three Peyronie patients and 162 healthy volunteers were included in the study. In both groups, demographic data, medical history, physical examination and erectile capacity were recorded. Serum 25‐hydroxyvitamin D, total cholesterol, low‐density lipoprotein, high‐density lipoprotein, triglyceride and testosterone levels were measured. The mean level of serum 25 (OH) D was significantly higher in men with Peyronie's disease compared with the controls (32.6 ± 7.9 ng/ml vs. 18.5 ± 6.6 ng/ml respectively. p < 0.001). There is a relationship between Peyronie's disease and high serum vitamin D levels. Also, increased low‐density lipoprotein and total cholesterol levels, diabetes mellitus, and cardiovascular diseases were associated with Peyronie's disease.     

Regeneration of pericardial tissue on absorbable polymer patches implanted into the pericardial sac. An immunohistochemical, ultrastructural and biochemical study in the sheep.

A new absorbable polymer prepared from polyhydroxybutyrate (PHB) was inserted as a pericardial patch in sheep to serve as a temporary scaffold for regeneration of pericardial tissue. Postoperative adhesions were rare or absent. The present study focuses on characterization of the regenerated surface cells. The luminal surface of the regenerated tissue was covered with a complete layer of mesothelium-like cells which at light and scanning electron microscopy resembled those in native pericardium. Immunohistochemical stainings for cytokeratin and thrombomodulin were positive in these cells. Heparan sulfate proteoglycan was found in a basement-membrane-like structure beneath the surface cells, as in the normal pericardium. Transmission electron microscopy of the regenerated surface revealed cells with the characteristics of mesothelium. Prostacyclin production in the regenerated tissue was similar to that in native pericardium. The results indicate regeneration of a mesothelial layer with many of the important functions of native mesothelial cells. This may explain the presently and previously observed prevention of pericardial adhesions after cardiac surgery in this field. Clinical testing of PHB patches as pericardial substitutes is warranted in cardiac surgery when pericardial closure is desired.     

Synergistic effect of low‐intensity pulsed ultrasound on growth factor stimulation of nucleus pulposus cells

Low‐intensity pulsed ultrasound (LIPUS) has been reported to stimulate the activity of various cells. We have reported that the capacity of human intervertebral nucleus pulposus cell line to synthesize proteoglycan (PG) was increased by exposure to LIPUS, and postulated that one of the mechanisms underlying this response was an increase in expression of the transforming growth factor‐β type I receptor gene (TGFβR1). Therefore, the present study was conducted to assess the synergistic effect of LIPUS and TGF‐β on nucleus pulposus cells harvested from canines. The cells were cultured under four different sets of conditions: control group (Group A), LIPUS group (Group B), TGF‐β1 group (Group C), and LIPUS + TGF‐β1 group (Group D). They were evaluated by measuring cell proliferation, PG synthesis, PG content, gene expression of TGFβR1, and TGF‐β1 concentration. There were no significant differences in proliferation during culture. However, PG synthesis and endogenous TGF‐β1 production increased and demonstrated a synergistic effect between LIPUS and TGF‐β. Because LIPUS is safe and noninvasive, the results of the present study suggest that it would be a promising new therapy for prevention of intervertebral disc degeneration, which is said to be one of the primary causes of low back pain. © 2007 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 25:1574–1581, 2007     

Transforming growth factor β1 regulates the expression of CCN2 in human keratinocytes via Smad‐ERK signalling

Connective tissue growth factor (CCN2/CTGF) and transforming growth factor β1 (TGF‐β1) are important regulators of skin wound healing, but controversy remains regarding their expression in epithelial cell lineages. Here, we investigate the expression of CCN2 in keratinocytes during reepithelialisation and its regulation by TGF‐β1. CCN2 was detected in the epidermis of healing full‐thickness porcine wounds. Human keratinocytes were incubated with or without 10 ng/ml TGF‐β1, and signalling pathways were blocked with 10‐μM SIS3 or 20‐μM PD98059. Semi‐quantitative real‐time PCR was used to study CCN2 mRNA expression, and western blot was used to measure CCN2, phosphorylated‐ERK1/2, ERK1/2, phosphorylated‐Smad3 and Smad2/3 proteins. CCN2 was transiently expressed in neoepidermis at the leading edge of the wound in vivo. In vitro, CCN2 expression was induced by TGF‐β1 at 2 hours (7·5 ± 1·9‐fold mRNA increase and 3·0 ± 0·6‐fold protein increase) and 12 hours (5·4 ± 1·9‐fold mRNA increase and 3·3 ± 0·6‐fold protein increase). Compared with inhibiting the SMAD pathway, inhibiting the mitogen‐activated protein kinase (MAPK) pathway was more effective in reducing TGF‐β1‐induced CCN2 mRNA and protein expression. Inhibition of the MAPK pathway had minimal impact on the activity of the SMAD pathway. CCN2 is expressed in keratinocytes in response to tissue injury or TGF‐β1. In addition, TGF‐β1 induces CCN2 expression in keratinocytes through the ras/MEK/ERK pathway. A complete understanding of CCN2 expression in keratinocytes is critical to developing novel therapies for wound healing and cutaneous malignancy.     

Donkey Pericardium as an Alternative Bioprosthetic Heart Valve Material

This study comparatively evaluates the characteristics of glutaraldehyde‐treated acellular bovine and donkey pericardium using histological and electronic microscopic observation techniques, shrinkage temperature, and mechanical properties, as well as determining calcium and phosphorus content at 4 and 8 weeks after the subcutaneous implantation of donkey and bovine pericardium in Wistar rats. Donkey pericardium was significantly thinner compared with bovine pericardium (1.622 ± 0.161 mm vs. 4.027 ± 0.401 mm, P < 0.0001) and was associated with significantly greater tensile strength (14.21 ± 3.81 MPa vs. 3.78 ± 1.20 MPa, P = 0.001) and elastic modulus (81.67 ± 20.41 MPa vs. 21.67 ± 11.69 MPa, P < 0.0001) over bovine pericardium. Shrinkage temperature of donkey pericardium was similar to that of bovine pericardium (87.43 ± 0.55°C vs. 87.50 ± 0.36°C, P = 0.810). No differences between groups were observed for maximum load (donkey: 21.64 ± 7.02 KN/m vs. bovine: 15.05 ± 4.50 KN/m, P = 0.082) and tear strength (donkey: 11.54 ± 5.33 MPa vs. bovine: 10.69 ± 3.77 MPa, P = 0.757). Calcium content was significantly lower in donkey pericardium compared with bovine pericardium at 4 weeks (690.15 ± 191.27 µg/g vs. 1381.73 ± 62.52 µg/g, P = 0.001) and 8 weeks (205.24 ± 62.40 µg/g vs. 910.48 ± 398.29 µg/g, P = 0.037). This preliminary study has confirmed that glutaraldehyde‐tanned donkey pericardium, demonstrating reduced calcification and increased tensile strength, may provide a suitable bioprosthetic valve substitute.     

Inflammation and Epithelial to Mesenchymal Transition in Lung Transplant Recipients: Role in Dysregulated Epithelial Wound Repair

Epithelial to mesenchymal transition (EMT) has been implicated in the pathogenesis of obliterative bronchiolitis (OB) after lung transplant. Although TNF‐α accentuates TGF‐β1 driven EMT in primary human bronchial epithelial cells (PBECs), we hypothesized that other acute pro‐inflammatory cytokines elevated in the airways of patients with OB may also accentuate EMT and contribute to dysregulated epithelial wound repair. PBECs from lung transplant recipients were stimulated with TGF‐β1 ± IL‐1β, IL‐8, TNF‐α or activated macrophages in co‐culture and EMT assessed. The quality and rate of wound closure in a standardized model of lung epithelial injury was assessed in response to above stimuli. Co‐treatment with TGF‐β1 + TNF‐α or IL‐1β significantly accentuates phenotypic and some functional features of EMT compared to TGF‐β1 alone. Co‐treatment with TGF‐β1 + TNF‐α or IL‐1β accelerates epithelial wound closure however the quality of repair is highly dysregulated. Co‐treatment with TGF‐β1 + IL‐8 has no significant effect on EMT or the speed or quality of wound healing. Activated macrophages dramatically accentuate TGF‐β1‐driven EMT and cause dysregulated wound repair. Crosstalk between macrophage‐derived acute inflammation in the airway and elevated TGF‐β1 may favor dysregulated airway epithelial repair and fibrosis in the lung allograft via EMT.     

Role of TGF‐β in a Mouse Model of High Turnover Renal Osteodystrophy

Altered bone turnover is a key pathologic feature of chronic kidney disease‐mineral and bone disorder (CKD‐MBD). Expression of TGF‐β1, a known regulator of bone turnover, is increased in bone biopsies from individuals with CKD. Similarly, TGF‐β1 mRNA and downstream signaling is increased in bones from jck mice, a model of high‐turnover renal osteodystrophy. A neutralizing anti‐TGF‐β antibody (1D11) was used to explore TGF‐β's role in renal osteodystrophy. 1D11 administration to jck significantly attenuated elevated serum osteocalcin and type I collagen C‐telopeptides. Histomorphometric analysis indicated that 1D11 administration increased bone volume and suppressed the elevated bone turnover in a dose‐dependent manner. These effects were associated with reductions in osteoblast and osteoclast surface areas. Micro‐computed tomography (µCT) confirmed the observed increase in trabecular bone volume and demonstrated improvements in trabecular architecture and increased cortical thickness. 1D11 administration was associated with significant reductions in expression of osteoblast marker genes (Runx2, alkaline phosphatase, osteocalcin) and the osteoclast marker gene, Trap5. Importantly, in this model, 1D11 did not improve kidney function or reduce serum parathyroid hormone (PTH) levels, indicating that 1D11 effects on bone are independent of changes in renal or parathyroid function. 1D11 also significantly attenuated high‐turnover bone disease in the adenine‐induced uremic rat model. Antibody administration was associated with a reduction in pSMAD2/SMAD2 in bone but not bone marrow as assessed by quantitative immunoblot analysis. Immunostaining revealed pSMAD staining in osteoblasts and osteocytes but not osteoclasts, suggesting 1D11 effects on osteoclasts may be indirect. Immunoblot and whole genome mRNA expression analysis confirmed our previous observation that repression of Wnt/β‐catenin expression in bone is correlated with increased osteoclast activity in jck mice and bone biopsies from CKD patients. Furthermore, our data suggest that elevated TGF‐β may contribute to the pathogenesis of high‐turnover disease partially through inhibition of β‐catenin signaling. © 2014 American Society for Bone and Mineral Research.     

Hyperactive Transforming Growth Factor‐β1 Signaling Potentiates Skeletal Defects in a Neurofibromatosis Type 1 Mouse Model

Dysregulated transforming growth factor beta (TGF‐β) signaling is associated with a spectrum of osseous defects as seen in Loeys‐Dietz syndrome, Marfan syndrome, and Camurati‐Engelmann disease. Intriguingly, neurofibromatosis type 1 (NF1) patients exhibit many of these characteristic skeletal features, including kyphoscoliosis, osteoporosis, tibial dysplasia, and pseudarthrosis; however, the molecular mechanisms mediating these phenotypes remain unclear. Here, we provide genetic and pharmacologic evidence that hyperactive TGF‐β1 signaling pivotally underpins osseous defects in Nf1^flox/?;Col2.3Cre mice, a model which closely recapitulates the skeletal abnormalities found in the human disease. Compared to controls, we show that serum TGF‐β1 levels are fivefold to sixfold increased both in Nf1^flox/?;Col2.3Cre mice and in a cohort of NF1 patients. Nf1‐deficient osteoblasts, the principal source of TGF‐β1 in bone, overexpress TGF‐β1 in a gene dosage–dependent fashion. Moreover, Nf1‐deficient osteoblasts and osteoclasts are hyperresponsive to TGF‐β1 stimulation, potentiating osteoclast bone resorptive activity while inhibiting osteoblast differentiation. These cellular phenotypes are further accompanied by p21‐Ras–dependent hyperactivation of the canonical TGF‐β1–Smad pathway. Reexpression of the human, full‐length neurofibromin guanosine triphosphatase (GTPase)‐activating protein (GAP)‐related domain (NF1 GRD) in primary Nf1‐deficient osteoblast progenitors, attenuated TGF‐β1 expression levels and reduced Smad phosphorylation in response to TGF‐β1 stimulation. As an in vivo proof of principle, we demonstrate that administration of the TGF‐β receptor 1 (TβRI) kinase inhibitor, SD‐208, can rescue bone mass deficits and prevent tibial fracture nonunion in Nf1^flox/?;Col2.3Cre mice. In sum, these data demonstrate a pivotal role for hyperactive TGF‐β1 signaling in the pathogenesis of NF1‐associated osteoporosis and pseudarthrosis, thus implicating the TGF‐β signaling pathway as a potential therapeutic target in the treatment of NF1 osseous defects that are refractory to current therapies. © 2013 American Society for Bone and Mineral Research.     

Sustained delivery of transforming growth factor beta three enhances tendon‐to‐bone healing in a rat model

Despite advances in surgical technique, rotator cuff repairs are plagued by a high rate of failure. This failure rate is in part due to poor tendon‐to‐bone healing; rather than regeneration of a fibrocartilaginous attachment, the repair is filled with disorganized fibrovascular (scar) tissue. Transforming growth factor beta 3 (TGF‐β3) has been implicated in fetal development and scarless fetal healing and, thus, exogenous addition of TGF‐β3 may enhance tendon‐to‐bone healing. We hypothesized that: TGF‐β3 could be released in a controlled manner using a heparin/fibrin‐based delivery system (HBDS); and delivery of TGF‐β3 at the healing tendon‐to‐bone insertion would lead to improvements in biomechanical properties compared to untreated controls. After demonstrating that the release kinetics of TGF‐β3 could be controlled using a HBDS in vitro, matrices were incorporated at the repaired supraspinatus tendon‐to‐bone insertions of rats. Animals were sacrificed at 14–56 days. Repaired insertions were assessed using histology (for inflammation, vascularity, and cell proliferation) and biomechanics (for structural and mechanical properties). TGF‐β3 treatment in vivo accelerated the healing process, with increases in inflammation, cellularity, vascularity, and cell proliferation at the early timepoints. Moreover, sustained delivery of TGF‐β3 to the healing tendon‐to‐bone insertion led to significant improvements in structural properties at 28 days and in material properties at 56 days compared to controls. We concluded that TGF‐β3 delivered at a sustained rate using a HBDS enhanced tendon‐to‐bone healing in a rat model. © 2011 Orthopaedic Research Society Published by Wiley Periodicals, Inc. J Orthop Res 29: 1099–1105, 2011     

Osteoblast‐Derived TGF‐β1 Stimulates IL‐8 Release Through AP‐1 and NF‐κB in Human Cancer Cells

Introduction: The bone marrow microenvironment is further enriched by growth factors released during osteoclastic bone resorption. It has been reported that the chemokine interleukin (IL)‐8 is a potent and direct activator of osteoclastic differentiation and bone resorption. However, the effect of bone‐derived growth factors on the IL‐8 production in human cancer cells and the promotion of osteoclastogenesis are largely unknown. The aim of this study was to investigate whether osteoblast‐derived TGF‐β1 is associated with osteolytic bone diseases. Materials and Methods: IL‐8 mRNA levels were measured using RT‐PCR analysis. MAPK phosphorylation was examined using the Western blot method. siRNA was used to inhibit the expression of TGF‐β1, BMP‐2, and IGF‐1. DNA affinity protein‐binding assay and chromatin immunoprecipitation assays were used to study in vitro and in vivo binding of c‐fos, c‐jun, p65, and p50 to the IL‐8 promoter. A transient transfection protocol was used to examine IL‐8, NF‐κB, and activator protein (AP)‐1 activity. Results: Osteoblast conditioned medium (OBCM) induced activation of IL‐8, AP‐1, and NF‐κB promoter in human cancer cells. Osteoblasts were transfected with TGF‐β1, BMP‐2, or IGF‐1 small interfering RNA, and the medium was collected after 48 h. TGF‐β1 but not BMP‐2 or IGF‐1 siRNA inhibited OBCM‐induced IL‐8 release in human cancer cells. In addition, TGF‐β1 also directly induced IL‐8 release in human cancer cells. Activation of AP‐1 and NF‐κB DNA‐protein binding and MAPKs after TGF‐β1 treatment was shown, and TGF‐β1–induced IL‐8 promoter activity was inhibited by the specific inhibitors of MAPK cascades. Conclusions: In this study, we provide evidence to show that the osteoblasts release growth factors, including TGF‐β1, BMP‐2, and IGF‐1. TGF‐β1 is the major contributor to the activation of extracellular signal‐related kinase (ERK), p38, and c‐Jun N‐terminal kinase (JNK), leading to the activation of AP‐1 and NF‐κB on the IL‐8 promoter and initiation of IL‐8 mRNA and protein release, thereby promoting osteoclastogenesis.     

Reduction of retrosternal and pericardial adhesions with rapidly resorbable polymer films.

BACKGROUND: The formation of postoperative cardiac adhesions makes a repeat sternotomy time consuming and dangerous. Many attempts have been made to solve this problem by using either drugs to inhibit fibrinolytic activity or different types of pericardial substitutes. The results have not been satisfactory. METHODS: The efficacy of bioresorbable film prototypes made of polyethylene glycol (EO) and polylactic acid (LA) (EO/LA = 1.5, 2.5, and 3.0) in the prevention of adhesions after cardiac operations in canine models was tested. After desiccation and abrasion of the epicardium, a transparent bioresorbable film was placed over the heart. The pericardium was closed to allow intrapericardial adhesions (n = 32) or left open and attached to the chest wall to induce retrosternal adhesions (n = 17). Postoperative recovery was similar among the groups. Retrosternal and pericardial adhesions were evaluated at necropsy 3 weeks later by assessing area, tenacity, and density of the adhesions. RESULTS: In the control dogs, tenacious, dense adhesions were observed. In contrast, adhesion formation was reduced at all sites covered by the films. The bioresorbable films were efficacious in the reduction of adhesion formation between epicardium and pericardium or between epicardium and sternum after cardiac operation. The EO/LA 1.5 film most effectively prevented the early adhesions. CONCLUSIONS: The bioresorbable films (EO/LA = 1.5, 2.5, and 3.0) significantly reduced adhesion formation, with EO/LA = 1.5 (Repel CV) being optimal. As the barrier was rapidly resorbed, the capsule formation induced by permanent barriers was avoided.