The Use of Biopolymers for Tissue Engineering
Creators: name: Vazquez-Portalatin, Nelda
Publisher: name: Purdue University Graduate School
Year: 2019
Subjects: Biomedical engineering not elsewhere classified
Regenerative medicine (incl. stem cells)
Agricultural biotechnology not elsewhere classified
Biochemistry and cell biology not elsewhere classified
Organic chemistry not elsewhere classified

description: Osteoarthritis (OA) is a degenerative joint disease characterized by cartilage damage and loss in the joints that affects approximately 27 million adults in the US. Tissue that is damaged by OA is a major health concern since cartilage tissue has a limited ability to self-repair due to the lack of vasculature in cartilage and low cell content. Tissue engineering efforts aim towards the development of cartilage repair strategies that mimic articular cartilage and are able to halt the progression of the disease as well as restore cartilage to its normal function.This study harnesses the biological activity of collagen type II, present in articular cartilage, and the superior mechanical properties of collagen type I by characterizing gels made of collagen type I and II blends (1:0, 3:1, 1:1, 1:3, and 0:1). The collagen blend hydrogels were able to incorporate both types of collagen and retain chondroitin sulfate (CS) and hyaluronic acid (HA). Cryoscanning electron microscopy images showed that the 3:1 ratio of collagen type I to type II gels had a lower void space percentage (36.4%) than the 1:1 gels (46.5%) and the complex modulus was larger for the 3:1 gels (G*=5.0 Pa) compared to the 1:1 gels (G*=1.2 Pa). The 3:1 blend consistently formed gels with superior mechanical properties compared to the other blends and has the potential to be implemented as a scaffold for articular cartilage engineering. Following the work done to characterize the collagen scaffolds, we studied whether an aggrecan mimic, CS-GAHb, composed of CS and HA binding peptides, GAH, and not its separate components, is able to prevent glycosaminoglycan (GAG) and collagen release when incorporated into chondrocyte-embedded collagen gels. Bovine chondrocytes were cultured and embedded in collagen type I scaffolds with CS, GAH, CS and GAH, or CS-GAHb molecules. Gels composed of 3:1 collagen type I and II with CS or CS-GAHb were also studied. The results obtained showed CS-GAHb is able to decrease GAG and collagen release and increase GAG retention in the gels. CS-GAHb also stimulated cytokine production during the initial days of scaffold culture. However, the addition of CS-GAHb into the chondrocyte-embedded collagen scaffolds did not affect ECM protein expression in the gels. The incorporation of collagen type II into the collagen type I scaffolds did not significantly affect GAG and cytokine production and ECM protein synthesis, but did increase collagen release. The results suggest the complex interaction between CS-GAHb, the chondrocytes, and the gel matrix make these scaffolds promising constructs for articular cartilage repair. Finally, we used Dunkin Hartley guinea pigs, a commonly used animal model of osteoarthritis, to determine if high frequency ultrasound can ensure intra-articular injections of the aggrecan mimic are accurately positioned in the knee joint. A high-resolution small animal ultrasound system with a 40 MHz transducer was used for image-guided injections. We assessed our ability to visualize important anatomical landmarks, the needle, and anatomical changes due to the injection. From the ultrasound images, we were able to visualize clearly the movement of anatomical landmarks in 75% of the injections. The majority of these showed separation of the fat pad (67.1%), suggesting the injections were correctly delivered in the joint space. The results demonstrate this image-guided technique can be used to visualize the location of an intra-articular injection in the joints of guinea pigs and we are able to effectively inject the aggrecan mimic into knee joints.All of the work presented here suggests that the addition of the aggrecan mimic to collagen I and collagen I and II scaffolds has shown that this type of construct could be useful for treating cartilage damage in the future.