Link to the University of Pittsburgh Homepage
Link to the University Library System Homepage Link to the Contact Us Form

CHARACTERIZATION OF SMOOTH MUSCLE CELL PHENOTYPE AND FUNCTIONALITY FOR POTENTIAL TISSUE ENGINEERING APPLICATIONS

Patel, Sanket N (2009) CHARACTERIZATION OF SMOOTH MUSCLE CELL PHENOTYPE AND FUNCTIONALITY FOR POTENTIAL TISSUE ENGINEERING APPLICATIONS. Master's Thesis, University of Pittsburgh. (Unpublished)

[img]
Preview
PDF
Primary Text

Download (2MB) | Preview

Abstract

Smooth muscle cell (SMC) embedded scaffolds have possible applications in treating diseased tissues that are rich in SMCs. Stress urinary incontinence (SUI) is an example of a disease that can be caused due to SMC dysfunction within the urinary sphincter. The goal of this thesis was to create a SMC-populated tissue engineered urethral wrap (TEUW) using autologous urethral SMCs (uSMCs), to be used as a cuff around the native urethra to integrate with the host tissue for providing mechanical and functional reinforcement to the diseased urethra. uSMCs were isolated from rat urethras. SMC phenotype was verified by immunofluorescence and western blotting. Isolation purity was assessed by staining uSMCs for skeletal muscle and urothelium markers since they are also present in the urethra. TEUWs were examined for SMC phenotype, apoptosis, mechanical and histological endpoints after culture.This thesis also evaluated the functionality of differentiated SMCs (dSMCs), which were derived via mechanical stimulation of bone marrow-derived mesenchymal stem cell (BMMSCs). The long-term objective is to use BMMSCs as an autologous source for SMCs in order to create TEUW-like tubular constructs for treating SMC related dysfunctions including, but not limited to SUI. uSMCs and dSMCs were assessed and compared for intracellular Ca2+ activity (fura-2) and contractile responses (live-cell) to various stimuli.Results of isolated uSMCs revealed expression of SMC markers and absence of skeletal and urothelium markers, suggesting isolation purity. uSMC-based TEUWs showed non-linear pressure-diameter profiles like soft tissues, greater compliance than the native urethra, and burst pressures similar to stem-cell based TEUWs. Both, uSMCs and dSMCs, exhibited intracellular Ca2+ activity, with and without extracellular Ca2+, vital for full SMC function. However, their failure to show morphological changes in the presence of agonists during contractility assessment indicated absence of mature SMCs. In summary, this study demonstrates proficient uSMC isolation, which represents an important step towards TEUW development, and that uSMCs and dSMCs are not fully functional at the differentiation stage tested. Future work should focus on increasing contractile protein expression by using matrix-like culture systems and/or biochemical stimulants. Following a systematic examination, SMC-populated TEUWs could be tested in an animal model.


Share

Citation/Export:
Social Networking:
Share |

Details

Item Type: University of Pittsburgh ETD
Status: Unpublished
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Patel, Sanket NSunnyNPatel@gmail.com
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Committee ChairVorp, David Avorpda@upmc.edu
Committee MemberDeasy, Bridget Mdeasybm@upmc.edu
Committee MemberSt.Croix, Claudette Mcls13@pitt.eduCLS13
Date: 25 September 2009
Date Type: Completion
Defense Date: 24 July 2009
Approval Date: 25 September 2009
Submission Date: 27 July 2009
Access Restriction: No restriction; Release the ETD for access worldwide immediately.
Institution: University of Pittsburgh
Schools and Programs: Swanson School of Engineering > Bioengineering
Degree: MSBeng - Master of Science in Bioengineering
Thesis Type: Master's Thesis
Refereed: Yes
Uncontrolled Keywords: smooth muscle; tissue engineering; urethra
Other ID: http://etd.library.pitt.edu/ETD/available/etd-07272009-083807/, etd-07272009-083807
Date Deposited: 10 Nov 2011 19:54
Last Modified: 15 Nov 2016 13:47
URI: http://d-scholarship.pitt.edu/id/eprint/8658

Metrics

Monthly Views for the past 3 years

Plum Analytics


Actions (login required)

View Item View Item