Buckenmeyer, Michael J.
(2020)
Bioengineering an Ovarian-specific ECM Hydrogel to Facilitate Intraovarian Follicle Delivery for Fertility Preservation.
Doctoral Dissertation, University of Pittsburgh.
(Unpublished)
Abstract
The ovarian reserve represents the absolute number of primordial follicles with the potential to release a fertilizable oocyte. A reduction of follicles occurs naturally with aging until the reserve is depleted and induces menopause. However, there is a growing population of post pubertal women who experience aberrant reproductive function prior to the onset of menopause, classified as premature ovarian failure (POF). A subset of POF cases directly result from surgical or chemical procedures used to ameliorate cancer or other ovarian-associated pathologies. The current gold standard used to treat iatrogenic POF is the transplantation of cryopreserved ovarian tissues, which has led to greater than 130 reported live-births to-date. This strategy has shown clinical success, but there remain risks of malignant cell transmission and complications due to surgery. We hypothesized that an ovarian-specific hydrogel could provide a minimally-invasive platform to facilitate the intraovarian delivery, engraftment and survival of isolated immature ovarian follicles.
The first aim of this study was to prepare a hydrogel from ovarian tissues that retained ovarian specificity. Porcine ovaries were decellularized using a series of enzymes and detergents to remove immunogenic components from the tissues. Decellularization significantly reduced dsDNA, while preserving ovarian tissue morphology, extracellular matrix (ECM) proteins and proteoglycans. Additionally, ovarian-specific hormones, such as anti-Müllerian hormone, estradiol and progesterone, were detected within decellularized ovaries. Ovarian ECM (OECM) hydrogels formed via physical crosslinking when exposed to physiological conditions. Hydrogel viscoelastic and ultrastructural properties were sensitive to changes in ECM concentration, while gelation time remained constant. Successful in vitro follicle maturation using OECM hydrogels demonstrated follicle compatibility.
The second aim of this study was to examine the efficacy of an OECM hydrogel as a vehicle to support intraovarian follicle delivery and graft survival within chemotherapy-induced POF (ciPOF) mice. Immature ovarian follicles from young DBA-GFP/nu female mice were enzymatically isolated and microinjected into the ovaries of ciPOF nude mice within the OECM hydrogel. Mating studies resulted in multiple litters containing pups derived from transplanted donor follicles. Transplanted ovarian tissues contained developing donor follicles suggesting hydrogel-facilitated intraovarian microinjection could offer a low-risk, minimally-invasive alternative for fertility preservation.
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Details
| Item Type: |
University of Pittsburgh ETD
|
| Status: |
Unpublished |
| Creators/Authors: |
| Creators | Email | Pitt Username | ORCID  |
|---|
| Buckenmeyer, Michael J. | mjb235@pitt.edu | mjb235 | |
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| ETD Committee: |
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| Date: |
29 September 2020 |
| Date Type: |
Publication |
| Defense Date: |
19 June 2020 |
| Approval Date: |
29 September 2020 |
| Submission Date: |
6 July 2020 |
| Access Restriction: |
1 year -- Restrict access to University of Pittsburgh for a period of 1 year. |
| Number of Pages: |
241 |
| Institution: |
University of Pittsburgh |
| Schools and Programs: |
Swanson School of Engineering > Bioengineering |
| Degree: |
PhD - Doctor of Philosophy |
| Thesis Type: |
Doctoral Dissertation |
| Refereed: |
Yes |
| Uncontrolled Keywords: |
Biomaterials, Extracellular Matrix, Hydrogels, Fertility Preservation |
| Date Deposited: |
29 Sep 2021 05:00 |
| Last Modified: |
29 Sep 2021 05:15 |
| URI: |
http://d-scholarship.pitt.edu/id/eprint/39308 |
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