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1.
J Surg Res ; 163(2): e105-12, 2010 Oct.
Article in English | MEDLINE | ID: mdl-20638677

ABSTRACT

BACKGROUND: Most research evaluating adipose-derived stem cells (ASC) uses tissue obtained from young, healthy patients undergoing plastic surgical procedures. Given the propensity of other adult stem cell lines to diminish with increasing patient age and co-morbidities, we assess the availability of ASC in elderly patients undergoing vascular surgical procedures, and evaluate their acquisition of endothelial cell (EC) traits to define their potential use in vascular tissue engineering. METHODS AND METHODS: Adipose tissue obtained by liposuction from patients undergoing vascular procedures (n = 50) was digested with collagenase and centrifuged to remove mature adipocytes. The resultant number of cells, defined as the stromal-vascular (SV) pellet, was quantified. Following a 7-d culture period and negative selection for CD31 and CD45, the resultant number of ASC was quantified. After culture in differentiating media (EMG-2), ASCs were tested for the acquisition of endothelial-specific traits (expression of CD31, realignment in shear, cord formation on Matrigel). RESULTS: The SV pellet contained 2.87 ± 0.34 × 10(5) cells/g fat, and the resultant number of ASCs obtained was 1.41 ± 0.18 × 10(5) cells/g fat. Flow cytometry revealed a homogeneous ASC population (>98% positive for CD13, 29, 90). Advanced age or co-morbidity (obesity, diabetes, renal or peripheral vascular disease) did not significantly alter yield of ASC. After culture in differentiating media (EMG-2), ASCs acquired each of the endothelial-specific traits. CONCLUSION: ASC isolation appears independent of age and co-morbidities, and ASCs harvested from patients with vascular disease retain their ability to differentiate into endothelial-like cells. Adipose tissue, therefore, is a practical source of autologous, adult stem cells for vascular tissue engineering.


Subject(s)
Adipose Tissue/cytology , Adult Stem Cells/cytology , Vascular Surgical Procedures , Adult , Age Factors , Aged , Cell Differentiation , Cell Separation , Comorbidity , Female , Humans , Male , Middle Aged , Tissue Engineering
2.
Tissue Eng Part A ; 16(1): 245-55, 2010 Jan.
Article in English | MEDLINE | ID: mdl-19698069

ABSTRACT

Use of adult adipose-derived stem cells (ASCs) as endothelial cell substitutes in vascular tissue engineering is attractive because of their availability. However, when seeded onto decellularized vascular scaffolding and exposed to physiological fluid shear force, ASCs are physically separated from the graft lumen. Herein we have investigated methods of increasing initial ASC attachment using luminal precoats and a novel protocol for the gradual introduction of shear stress to optimize ASC retention. Fibronectin coating of the graft lumen increased ASC attachment by nearly sixfold compared with negative controls. Gradual introduction of near physiological fluid shear stress using a novel bioreactor whereby flow rate was increased every second at a rate of 1.5 dynes/cm(2) per day resulted in complete luminal coverage compared with near complete cell loss following conventional daily abrupt increases. An upregulation of the alpha(5)beta(1) integrin was evinced following exposure to shear stress, which accounts for the observed increase in ASC retention on the graft lumen. These results indicated a novel method for seeding, conditioning, and retaining of adult stem cells on a decellularized vein scaffold within a high-shear stress microenvironment.


Subject(s)
Adipose Tissue/metabolism , Adult Stem Cells/metabolism , Integrin alpha5beta1/biosynthesis , Saphenous Vein , Stress, Physiological , Up-Regulation , Adipose Tissue/cytology , Adult Stem Cells/cytology , Bioreactors , Cell Adhesion , Cell Culture Techniques , Humans
3.
Ann Surg ; 246(3): 472-7; discussion 477-80, 2007 Sep.
Article in English | MEDLINE | ID: mdl-17717451

ABSTRACT

OBJECTIVE: Technological advances in vascular surgery have changed the field dramatically over the past 10 years. Herein, we evaluate the impact of endovascular procedures on general surgery training. METHODS: National operative data from the Residency Review Committee for Surgery were examined from 1997 through 2006. Total major vascular operations, traditional open vascular operations and endovascular procedures were evaluated for mean number of cases per graduating chief general surgery resident (GSR) and vascular surgery fellow (VSF). RESULTS: As endovascular surgical therapies became widespread, GSR vascular case volume decreased 34% over 10 years, but VSF total cases increased 78%. GSR experience in open vascular operations decreased significantly, as evidenced by a 52% decrease (P < 0.0001) in elective open AAA repair. VSFs have also seen significant decreases in open vascular procedures. Experience in endovascular procedures has increased for both general surgery and vascular residents, but the increase has been much larger in absolute number for VSFs. CONCLUSIONS: GSR experience in open vascular procedures has significantly decreased as technology has advanced within the field. Unlike VSFs, this loss has not been replaced by direct experience with endovascular training. These data demonstrate the impact technology can have on how we currently train general surgeons. New educational paradigms may be necessary in which either vascular surgery as an essential component is abandoned or training in catheter-based interventions becomes required.


Subject(s)
Education, Medical, Graduate/standards , General Surgery/education , Specialties, Surgical/education , Specialties, Surgical/standards , Vascular Surgical Procedures/education , Analysis of Variance , Female , Humans , Internship and Residency/standards , Male , United States
4.
J Shoulder Elbow Surg ; 14(1 Suppl S): 79S-83S, 2005.
Article in English | MEDLINE | ID: mdl-15726091

ABSTRACT

Shoulder overuse injuries, especially those to the supraspinatus tendon of the rotator cuff, are common musculoskeletal disorders. Unfortunately, little is known about the disease etiology and pathogenesis. The objective of this study was to determine the levels of specific inflammatory and angiogenic markers in a rat supraspinatus tendon overuse injury model. We hypothesized that inflammation would not be present early in the overuse protocol. Conversely, we hypothesized that the overuse protocol would result in increased angiogenesis early. Increases in five-lipoxygenase activating protein, cyclooxygenase-2, vascular endothelial growth factor, and von Willebrand factor were evaluated by use of reverse transcription-polymerase chain reaction from 1 day through 16 weeks of treadmill running (overuse protocol). These results provide important information on the role of angiogenesis and inflammation in the disease process. Future studies will further evaluate the mechanisms of the disease process as well as potential targeted treatment modalities.


Subject(s)
Cumulative Trauma Disorders/metabolism , Rotator Cuff Injuries , Rotator Cuff/metabolism , 5-Lipoxygenase-Activating Proteins , Animals , Carrier Proteins/biosynthesis , Carrier Proteins/genetics , Cumulative Trauma Disorders/complications , Cyclooxygenase 2 , Disease Models, Animal , Inflammation/etiology , Membrane Proteins/biosynthesis , Membrane Proteins/genetics , Neovascularization, Pathologic/etiology , Prostaglandin-Endoperoxide Synthases/biosynthesis , Prostaglandin-Endoperoxide Synthases/genetics , RNA, Messenger/biosynthesis , Rats , Rats, Sprague-Dawley , Vascular Endothelial Growth Factors/biosynthesis , Vascular Endothelial Growth Factors/genetics , von Willebrand Factor
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