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Show SCHOOL OF MEDICINE AND HEALTH SCIENCES UNDERGRADUATE RESEARCH ABSTRACTS COLLAGEN REGENERATION USING MESENCHYMAL STEM CELLS AND EXTRACELLULAR MATRIX Amber Staples (Dolly Holt, Francisco Silva, Don Atkinson, Amit N. Patel) Department of Surgery University of Utah Healthy joint cartilage extracellular matrix (ECM) consists of collagen Type I, Type II, and proteoglycans secreted by chondrocytes.-" Due to low vascularization, isolation, and decreasing activity with age, chondrocytes do not substantially regenerate cartilage. Thus, the body may not be able to adequately correct cartilagenous injuries such as those found in osteoarthritic joints. The purpose of this study was to determine if cartilage regeneration could be improved by the addition of increased nutrients, growth factors, and exogenous ECM. W e chose to use human platelet lysate (HPL) to add exogenous growth factors and nutrients and human adipose ECM as a rich source of collagens, elastins, and proteoglycans. The effectiveness of HPL and ECM in upregulating collagen production was tested in two-dimensions (2D) using 6-well plates and in three-dimensions (3D) with cell pellets. Bone-marrow-derived mesenchymal stem cells were passaged into 6 well plates and centrifuged into cell pellets in four conditions: chondrocyte differentiation media (CDM), C D M plus HPL, C D M plus ECM, and C D M plus HPL and ECM. When grown in 2D, chondrocytes spontaneously delaminated and formed balls after 12 days, a direct result of cells differentiating into chondrocytes and producing collagen. Cells in C D M plus HPL and ECM balled up first, followed by cells in C D M plus HPL, and finally those in C D M plus ECM. Cells in CDM alone never balled up. These results suggest the cells with C D M plus HPL and E C M upregulated cellular differentiation and E C M deposition more than any other group. Preliminary testing for RNA in the cells grown in 2 D showed maximum RNA quantities in the cells with C D M plus HPL and ECM, indicating increased cellular proliferation. Furthermore, staining of cells grown in 2 D showed large quantities of sulfated substances, substantiating the differentiation of the stem cells into chondrocytes. PCR testing will be performed to determine if the cells in C D M plus HPL and E C M produced more collagen (Type I and Type II) than the other groups. These results would be significant because, with further testing, an injectable gel containing HPL and E C M could be developed that could significantly improve current treatments for osteoarthritis by facilitating increased cartilage regeneration and potentially eliminating the need for surgery. Amber Staples Amit N Patel 131 |