Biomedical subjects

Julie R Fuchs

Publications and source records attributed to Julie R Fuchs.

17 recordsLinked to original sources

PubMed2006

Diaphragmatic repair through fetal tissue engineering: a comparison between mesenchymal amniocyte- and myoblast-based constructs.

PURPOSE: We have previously shown that fetal tissue engineering is a preferred alternative to diaphragmatic repair in a large animal model. This study was aimed at comparing diaphragmatic constructs seeded with mesenchymal amniocytes and fetal myoblasts in this model. METHODS: Neonatal lambs (n = 14) underwent repair of an experimental diaphragmatic defect with identical scaffolds, either seeded with labeled autologous cells (mesenchymal amniocytes in group 1 and fetal myoblasts in group 2) or as an acellular graft (group 3). At 1 to 12 months postoperatively, implants were harvested for multiple analyses. RESULTS: Repair failure (reherniation or eventration) was significantly higher in group 3 than in groups 1 and 2, with no difference between groups 1 and 2. Seeded fetal myoblasts quickly lost their myogenic phenotype in vivo. All grafts contained cells with a fibroblastic-myofibroblastic profile. Elastin concentrations and both modular and ultimate tensile strengths were significantly higher in group 1 than in groups 2 and 3. There were no differences in glycosaminoglycans and type I collagen levels among the groups. CONCLUSIONS: Diaphragmatic repair with a mesenchymal amniocyte-based engineered tendon leads to improved structural outcomes when compared with equivalent fetal myoblast-based and acellular grafts. The amniotic fluid is a preferred cell source for tissue-engineered diaphragmatic reconstruction.

Animals↗

PubMed2006

Postnatal myocardial augmentation with skeletal myoblast-based fetal tissue engineering.

BACKGROUND: Cardiac anomalies constitute the most common birth defects, many of which involve variable myocardial deficiencies. Therapeutic options for structural myocardial repair remain limited in the neonatal population. This study was aimed at determining whether engineered fetal muscle constructs undergo milieu-dependent transdifferentiation after cardiac implantation, thus becoming a potential means to increase/support myocardial mass after birth. METHODS: Myoblasts were isolated from skeletal muscle specimens harvested from fetal lambs, labeled by transduction with a retrovirus-expressing green fluorescent protein, expanded in vitro, and then seeded onto collagen hydrogels. After birth, animals underwent autologous implantation of the engineered constructs (n = 8) onto the myocardium as an onlay patch. Between 4 and 30 weeks postoperatively, implants were harvested for multiple analyses. RESULTS: Fetal and postnatal survival rates were 89% and 100%, respectively. Labeled cells were identified within the implants at all time points by immunohistochemical staining for green fluorescent protein. At 24 and 30 weeks postimplantation, donor cells double-stained for green fluorescent protein and Troponin I, while losing skeletal (type II) myosin expression. CONCLUSIONS: Fetal skeletal myoblasts engraft in native myocardium up to 30 weeks after postnatal, autologous implantation as components of engineered onlay patches. These cells also display evidence of time-dependent transdifferentiation toward a cardiomyocyte-like lineage. Further analysis of fetal skeletal myoblast-based constructs for the repair of congenital myocardial defects is warranted.

Animals↗

PubMed2006

Fetal hepatic haematopoiesis is modulated by arterial blood flow to the liver.

We describe an as yet unrecognised relationship between fetal hepatic haematopoiesis and arterial blood flow to the liver. To increase hepatic arterial flow, the common bile duct (CBD) was ligated in fetal lambs. Reduction of hepatic arterial flow was accomplished in age-matched animals by hepatic artery (HA) ligation. Multiple analyses performed before term showed a significant increase in haematopoietic cell density in CBD animals when compared with sham controls and HA animals. In contrast, HA animals demonstrated a decrease in liver haematopoietic activity. Fetal hepatic haematopoiesis is dependent upon arterial blood flow to the liver.

Animals↗

PubMed2005

Hydrogel optimization for cultured elastic chondrocytes seeded onto a polyglycolic acid scaffold.

The purpose of this study was to compare the effect of different hydrogels on the production of tissue-engineered cartilage based on polyglycolic acid (PGA). Chondrocytes were isolated from adult sheep auricles. Alginate, Type I collagen, methylcellulose, and pluronic F127 hydrogels were evaluated, as were controls prepared without hydrogels. Proliferated chondrocytes were mixed with each hydrogel at 20 x 10(6) cells/mL and seeded onto PGA (1 x 1 x 0.2 cm, n = 60). The constructs were cultured with serum-free medium containing 5 ng/mL TGF-beta(2) and 5 ng/mL des(1-3)IGF-I in rotational bioreactors for up to 6 weeks. The cellular morphology, histology, and biochemistry were analyzed. Type I collagen, methylcellulose, and pluronic F127 displayed improved cartilage matrix deposition in terms of histology and biochemistry compared to alginate. It was not concluded that the combined seeding of chondrocytes and hydrogels on a PGA scaffold had significantly better effects than cell seeding without hydrogels. However, the histology and other useful findings in this ECM analyses suggested that Type I collagen and MC hydrogels were the best candidates for cartilage regeneration, because of their stimulation for chondrocyte proliferation in a three-dimensional culture as well as cartilage regeneration.

Animals↗

PubMed2005

Intratracheal pulmonary ventilation improves gas exchange during laparoscopy in a pediatric lung injury model.

BACKGROUND/PURPOSE: This study was aimed at determining whether intraoperative intratracheal pulmonary ventilation (ITPV) could prevent/treat respiratory complications of laparoscopy in a model of pediatric pulmonary insufficiency. METHODS: Severe lung injury was induced in 0- to 2-month-old lambs (n = 5) by endotracheal saline lavage. Animals then underwent establishment of CO2 pneumoperitoneum. Intraperitoneal pressures were progressively raised from 0 to 15 mm Hg, at intervals of 5 mm Hg. At each interval, blood gas and hemodynamic data were recorded, 20 minutes after initiation of both conventional ventilation and pure ITPV. All ventilatory parameters were constant and identical on both modes of ventilation. RESULTS: On conventional ventilation, severe respiratory acidosis and hypoxemia ensued at intraperitoneal pressures of 5 mm Hg and 10 mm Hg or more, respectively. Compared with conventional ventilation, ITPV led to statistically significant decreases in PCO2 at intraperitoneal pressures of 5 mm Hg (43.2 +/- 5.2 vs 56.1 +/- 6.6 mm Hg) and 10 mm Hg (45.1 +/- 3.2 vs 61 +/- 6.3 mm Hg) and to significant increases in PO2 at 10 mm Hg (92 +/- 10.2 vs 61 +/- 8.1 mm Hg), resolving the acidosis and hypoxemia at those pressure levels. CONCLUSIONS: Compared with conventional ventilation, ITPV improves both CO2 removal and oxygenation during CO2 pneumoperitoneum in a pediatric lung injury model. Intratracheal pulmonary ventilation may be a safer intraoperative mode of ventilation for neonates and children with respiratory failure who require laparoscopy.

Acidosis, Respiratory↗

PubMed2005

Cost considerations and applicant characteristics for the Pediatric Surgery Match.

BACKGROUND/PURPOSE: Formal training in pediatric surgery is highly competitive. The limited number of accredited positions has historically favored applicants with basic science experience, numerous publications, national presentations, and exposure to well-known pediatric surgeons. This review analyzes characteristics of successful applicants and cost associated with the Match. METHODS: A survey was e-mailed to 45 applicants after the 2003 Match. Geographic provenance, demographics, qualifications, costs, and valued program characteristics were assessed. Statistics were formulated by chi2 and Student's t test. RESULTS: Thirty-six applicants (80%) responded. Successful characteristics for matched vs unmatched included number of publications, 11.2 vs 5.7 (P < .01); first-author designation, 6.4 vs 3.1 (P = .02); basic science papers, 5.7 vs 1.7 (P < .01); national presentations, 5.8 vs 2.4 (P = .02); and presentations at pediatric surgical meetings, 2.0 vs 0.6 (P = .04). Ninety percent of matched applicants took time off to perform basic science research (P < .01). Average candidate expense was $6974, which represented 14% of pretax salary. Forty-one percent of applicants noted that cost limited the number of interviews taken. Fifty percent preferred a regional interview process to limit expense. Candidates ranked case diversity, volume, and mentor's advice as the most valued program characteristics. Successful applicants matched at their fifth rank on average. Eighty-six percent of unsuccessful applicants will reapply. CONCLUSIONS: Results of this study are important to those interested in the future of pediatric surgery. Successful applicants were shown to have several national presentations and multiple scientific publications, especially in basic sciences. Applicant costs are high, totaling more than $236,000 for survey respondents.

Adult↗

PubMed2005

In vitro cartilage regeneration from proliferated adult elastic chondrocytes.

The purpose of this study was to investigate cellular feasibility in the proliferation and differentiation status of adult chondrocytes for cartilage regeneration in comparison to fetal chondrocytes. Primary cells were isolated from adult (n = 6) and fetal (n = 6) sheep ear cartilages and expanded in 10% fetal bovine serum (FBS) containing Ham's F12 medium, in which adult and fetal cell proliferation rates were compared using a WST-1 assay kit. Approximately 4 million cells were seeded onto each 1 x 1 x 0.2-cm (200 microL) nonwoven fabric scaffold made from polyglycolic acid. Cell/polymer constructs were cultured in serum-free DMEM/F12 medium supplemented with 5 ng/mL TGF-beta2 and 5 ng/mL des(1-3)IGF-I (adult chondrocytes, group A) or in 10% FBS containing Ham's F12 medium (adult chondrocytes, group B, and fetal chondrocytes, group C) as controls in a rotating bioreactor for 6 weeks. The proliferation assay showed that fetal cells had a significantly better growth potential than did adult cells. Histology and extracellular matrix analyses revealed that groups A and C qualitatively displayed better matrix deposition than did group B. In conclusion, although adult sheep elastic chondrocytes had less growth potential than did fetal cells, the serum-free medium supplemented with growth factors significantly enhanced the production of cartilage matrix secreted from proliferated adult sheep elastic chondrocytes.

Age Factors↗

PubMed2005

Cartilage engineering from ovine umbilical cord blood mesenchymal progenitor cells.

We aimed to determine whether three-dimensional (3D) cartilage could be engineered from umbilical cord blood (CB) cells and compare it with both engineered fetal cartilage and native tissue. Ovine mesenchymal progenitor cells were isolated from CB samples (n=4) harvested at 80-120 days of gestation by low-density fractionation, expanded, and seeded onto polyglycolic acid scaffolds. Constructs (n=28) were maintained in a rotating bioreactor with serum-free medium supplemented with transforming growth factor-beta1 for 4-12 weeks. Similar constructs seeded with fetal chondrocytes (n=13) were cultured in parallel for 8 weeks. All specimens were analyzed and compared with native fetal cartilage samples (n=10). Statistical analysis was by analysis of variance and Student's t-test (p<.01). At 12 weeks, CB constructs exhibited chondrogenic differentiation by both standard and matrix-specific staining. In the CB constructs, there was a significant time-dependent increase in extracellular matrix levels of glycosaminoglycans (GAGs) and type-II collagen (C-II) but not of elastin (EL). Fetal chondrocyte and CB constructs had similar GAG and C-II contents, but CB constructs had less EL. Compared with both hyaline and elastic native fetal cartilage, C-II and EL levels were, respectively, similar and lower in the CB constructs, which had correspondingly lower and similar GAG levels than native hyaline and elastic fetal cartilage. We conclude that CB mesenchymal progenitor cells can be successfully used for the engineering of 3D cartilaginous tissue in vitro, displaying select histological and functional properties of both native and engineered fetal cartilage. Cartilage engineered from CB may prove useful for the treatment of select congenital anomalies.

Animals↗

PubMed2004

Massive iliac artery bleeding in patients receiving intraoperative radiation therapy for advanced rectal cancer: report of two cases.

Multimodality treatment for advanced rectal cancer, including intraoperative radiation therapy, is associated with significant complications. Unfortunately, because of the high local recurrence rates and mortality in these patients, the treatment regimen of external beam radiation therapy, chemotherapy, extensive resection, and intraoperative radiation therapy is one of the few potentially curative options available. Here we report two cases of massive iliac artery bleeding after multimodality treatment for advanced rectal cancer, which included intraoperative radiation therapy.

Hemorrhage↗

PubMed2004

Management of spontaneous colonic perforation in Ehlers-Danlos syndrome type IV.

A 14-year-old girl with a family history of fatal colonic rupture, presented with a 2-day history of abdominal pain and signs of peritonitis. At laparotomy, a full-thickness perforation of the sigmoid colon was found, which was exteriorized as a loop colostomy. Subsequently, molecular studies of the patient's cultured fibroblasts found a point mutation in the COL3A1 gene, confirming a diagnosis of Ehlers-Danlos syndrome type IV (EDS-IV). Four and a half years later, a total abdominal colectomy and ileoproctostomy were performed, restoring intestinal continuity. At 5 years follow-up, the patient has had no further complications. Although spontaneous colonic perforation is a well-reported manifestation of EDS-IV, a consensus on the surgical management of this complication in EDS-IV has yet to be determined. Given the high rate of reperforation in EDS-IV when the colon is left in place and the low incidence of reported small bowel and rectal perforations, subtotal colectomy is a reasonable treatment. Primary anastomosis and avoidance of an end-ileostomy was possible in this young patient, with no evidence of anastomotic leakage nor reperforation to date. Lifelong close follow-up should be continued in these patients, because the natural history of this anatomy in EDS-IV is not known.

Abdominal Pain↗

PubMed2004

Diaphragmatic reconstruction with autologous tendon engineered from mesenchymal amniocytes.

PURPOSE: This study examined the effects of amniocyte-based engineered tendons on partial diaphragmatic replacement. METHODS: Ovine mesenchymal amniocytes were labeled with green fluorescent protein (GFP), expanded, and seeded into a collagen hydrogel. Composite grafts (20 to 25 cm2) based on acellular dermis (group I), or acellular small intestinal submucosa (group II) received either a cell-seeded or an acellular hydrogel within their layers. Newborn lambs (n = 20) underwent partial diaphragmatic replacement with either an acellular or a cellular autologous construct from either group. At 3 to 12 months' postoperatively, implants were subjected to multiple analyses. RESULTS: Diaphragmatic hernia recurrence was significantly higher in animals with acellular grafts (5 of 5) then in animals with cellular ones (1 of 4) in group I (P <.05) but not in group II (3 of 6 and 4 of 5, respectively). Cellular grafts had higher modular (5.27 +/- 1.98 v. 1.27 +/- 0.38 MPa) and ultimate (1.94 +/- 0.70 v. 0.29 +/- 0.05 MPa) tensile strength than acellular implants in group I (P <.05), but not in group II. Quantitative analyses showed no differences in extracellular matrix components between cellular and acellular implants in either group. All cellular implants showed GFP-positive cells. CONCLUSIONS: Diaphragmatic repair with an autologous tendon engineered from mesenchymal amniocytes leads to improved mechanical and functional outcomes when compared with an equivalent acellular bioprosthetic repair, depending on scaffold composition. The amniotic fluid may be a preferred cell source for engineered diaphragmatic reconstruction.

Amniotic Fluid↗

PubMed2003

Fetal tracheal augmentation with cartilage engineered from bone marrow-derived mesenchymal progenitor cells.

BACKGROUND/PURPOSE: The authors have described previously the use of engineered fetal cartilage in a large animal model of fetal tracheal repair. This study was aimed at comparing cartilage engineered from bone marrow-derived stromal cells (BMSC) to native and engineered cartilage, in this model. METHODS: Ovine BMSC were expanded in vitro, seeded onto biodegradable scaffolds, and maintained in transforming growth factor beta 1 (TGF-beta1)-supplemented medium for 3 months (group I). Identical scaffolds were seeded with fetal chondrocytes (group II). All constructs were analyzed in vitro, implanted into fetal tracheas, and harvested after birth for further analysis. RESULTS: There were no differences in survival between the groups. All BMSC-based constructs exhibited chondrogenic differentiation. Matrix analyses in vitro showed that both groups had similar levels of glycosaminoglycans (GAG) and type II collagen (C-II), but lower levels of elastin when compared with native fetal cartilage. Yet, compared with group II, group I had higher levels of GAG, equal levels of C-II, and lower levels of elastin. However, remodeling resulted in no differences between the 2 groups in any of these variables in vivo. CONCLUSIONS: The bone marrow may be a useful cell source for cartilage engineering aimed at the surgical repair of severe congenital tracheal anomalies, such as tracheal atresia and agenesis, in utero.

Animals↗

PubMed2003

Fetal tissue engineering: chest wall reconstruction.

BACKGROUND/PURPOSE: This study was aimed at applying fetal tissue engineering to chest wall reconstruction. METHODS: Fetal lambs underwent harvest of elastic and hyaline cartilage specimens. Once expanded in vitro, fetal chondrocytes were seeded onto synthetic scaffolds, which then were placed in a bioreactor. After birth, fetal cartilage constructs (n = 10) were implanted in autologous fashion into the ribs of all lambs (n = 6) along with identical, but acellular scaffolds, as controls (n = 6). Engineered and acellular specimens were harvested for analysis at 4 to 12 weeks postimplantation. Standard histology and matrix-specific staining were performed both before implantation and after harvest on all constructs. RESULTS: Regardless of the source of chondrocytes, all fetal constructs resembled hyaline cartilage, both grossly and histologically, in vitro. In vivo, engineered implants retained hyaline characteristics for up to 10 weeks after implantation but remodeled into fibrocartilage by 12 weeks postoperatively. Mononuclear inflammatory infiltrates surrounding residual PGA/PLLA polymer fibers were noted in all specimens but most prominently in the acellular controls. CONCLUSIONS: Engineered fetal cartilage can provide structural replacement for at least up to 10 weeks after autologous, postnatal implantation in the chest wall. Fetal tissue engineering may prove useful for the treatment of severe congenital chest wall defects at birth.

Animals↗

PubMed2003

Fetal tissue engineering: in vitro analysis of muscle constructs.

BACKGROUND/PURPOSE: This study was aimed at examining the impact of different tissue engineering techniques on fetal muscle construct architecture. METHODS: Myoblasts from ovine specimens of fetal skeletal muscle were expanded in culture and their growth rates determined. Cells were seeded at different densities onto 3 scaffold types, namely polyglycolic acid (PGA) treated with poly-l-lactic acid (PLLA), a composite of PGA with poly-4-hydroxybutyrate (P4HB), and a collagen hydrogel. Constructs were maintained in a bioreactor and submitted to histologic, scanning electron microscopy, and DNA analyses at different time-points. Statistical analysis was by the likelihood ratio and paired Student's t tests (P <.05). RESULTS: Fetal myoblasts proliferated at faster rates than expected from neonatal cells. Cell attachment was enhanced in the PGA/PLLA matrix and collagen hydrogel when compared with the PGA/P4HB composite. Necrosis was observed at the center of all constructs, directly proportional to cell seeding density and time in the bioreactor. CONCLUSIONS: Fetal myoblasts can be expanded rapidly in culture and attach well to PGA/PLLA, as well as collagen hydrogel but less optimally to PGA/P4HB. Excessive cell seeding density and bioreactor time may worsen final construct architecture. These findings should be considered during in vivo trials of muscle replacement by engineered fetal constructs.

Animals↗

PubMed2002

Impact of abdominal CT imaging on the management of appendicitis: an update.

BACKGROUND: Abdominal computed tomographic scanning (ACTS) has recently been shown to be an accurate diagnostic tool for appendicitis and may improve the negative exploration rate in our patient population. MATERIALS AND METHODS: We reviewed 224 patients evaluated for appendicitis during 1998. Forty-two patients underwent appendectomy on clinical grounds alone (Group I), 182 patients underwent ACTS (Group II), and 79 patients in Group II were explored for appendicitis. Diagnostic errors, alternative diagnoses, and perforation rates were noted. RESULTS: There were five negative explorations in Group I (11.9%) and five in Group II (6.3%), resulting in a combined negative rate of 8.3%. The negative exploration rate in women was 23.5% in Group I and 5.3% in Group II (P = 0.07), producing a combined negative rate of 10.9%. Fifty-eight alternative diagnoses were made by ACTS. The ACTS made a critical difference in the management of 67% of patients over 50 years of age and in 79% of Group II patients. CONCLUSIONS: The negative exploration rate for appendicitis at our institution fell from 13.6 to 8.3% with selective use of ACTS. The most striking benefit occurred in women and in patients over 50 years of age.

Abdomen↗

PubMed2002

Fetal tissue engineering: in utero tracheal augmentation in an ovine model.

BACKGROUND/PURPOSE: This study was aimed at comparing fetal tissue engineering with autologous free grafting in an ovine model of in utero tracheal repair. METHODS: Chondrocytes were isolated from both elastic and hyaline cartilage specimens harvested from fetal lambs and expanded in vitro. Cells were seeded dynamically onto biodegradable scaffolds, which then were maintained in a rotating bioreactor for 6 to 8 weeks. Constructs subsequently were implanted into fetal tracheas (n = 15), in a heterologous fashion (group I). In group II, fetuses (n = 5) received autologous free grafts of elastic cartilage harvested from the ear as tracheal implants. In vivo specimens were harvested for histologic analysis at different time-points postimplantation. RESULTS: In the 12 of 15 surviving fetuses of group I, all constructs were found to resemble normal hyaline cartilage, engraft well despite their heterologous origin, and display time-dependent epithelialization derived from the native trachea. All autologous free grafts were engrafted and epithelialized at birth, retaining histologic characteristics of elastic cartilage, but were more deformed than engineered constructs. Of the lambs allowed to reach term, 5 of 5 in the engineered group and 4 of 5 in the free graft group could breathe spontaneously. CONCLUSIONS: (1) Tissue-engineered cartilage, as well as autologous free grafts, can be implanted successfully into the fetal trachea, resulting in engraftment and function. (2) Engineered cartilage provides enhanced structural support after implantation into the fetal trachea when compared with free grafts. Prenatal tracheoplasty may prove useful for the treatment of severe congenital tracheal malformations.

Animals↗

PubMed2002

Engineered fetal cartilage: structural and functional analysis in vitro.

BACKGROUND/PURPOSE: This study was aimed at characterizing the structure and function of engineered fetal cartilage in vitro. METHODS: Chondrocytes from ovine specimens of fetal elastic, fetal hyaline, and adult elastic cartilage were expanded in culture and their growth rates determined. Cells were seeded onto synthetic scaffolds, which were then maintained in a bioreactor. Matrix deposition was determined by specific staining and quantitative assays for glycosaminoglycans (GAG), type II collagen (CII), and elastin, as well as compared with native tissue. Statistical analysis was by analysis of variance (ANOVA) and Students' t test, with significance set at P less than.01. RESULTS: Fetal elastic chondrocytes grew significantly faster than all other cell types. All fetal constructs resembled hyaline cartilage, regardless of the cell source. There were significantly higher levels of GAG and CII in fetal versus adult constructs, but no significant difference between fetal constructs from different sources. Unlike their adult counterparts, fetal constructs had GAG and CII levels similar to native tissues. CONCLUSIONS: Fetal chondrocytes can be rapidly expanded in culture. Compared with adult constructs, matrix deposition is enhanced in engineered fetal cartilage, which closely resembles native tissue, regardless of the cell source. Engineered fetal cartilage may be a preferable option during surgical reconstruction of select congenital anomalies.

Analysis of Variance↗