Research Article: Enhanced articular cartilage by human mesenchymal stem cells in enzymatically mediated transiently RGDS-functionalized collagen-mimetic hydrogels

Date Published: March 15, 2017

Publisher: Elsevier

Author(s): Paresh A. Parmar, Jean-Philippe St-Pierre, Lesley W. Chow, Christopher D. Spicer, Violet Stoichevska, Yong Y. Peng, Jerome A. Werkmeister, John A.M. Ramshaw, Molly M. Stevens.


Recapitulation of the articular cartilage microenvironment for regenerative medicine applications faces significant challenges due to the complex and dynamic biochemical and biomechanical nature of native tissue. Towards the goal of biomaterial designs that enable the temporal presentation of bioactive sequences, recombinant bacterial collagens such as Streptococcal collagen-like 2 (Scl2) proteins can be employed to incorporate multiple specific bioactive and biodegradable peptide motifs into a single construct. Here, we first modified the backbone of Scl2 with glycosaminoglycan-binding peptides and cross-linked the modified Scl2 into hydrogels via matrix metalloproteinase 7 (MMP7)-cleavable or non-cleavable scrambled peptides. The cross-linkers were further functionalized with a tethered RGDS peptide creating a system whereby the release from an MMP7-cleavable hydrogel could be compared to a system where release is not possible. The release of the RGDS peptide from the degradable hydrogels led to significantly enhanced expression of collagen type II (3.9-fold increase), aggrecan (7.6-fold increase), and SOX9 (5.2-fold increase) by human mesenchymal stem cells (hMSCs) undergoing chondrogenesis, as well as greater extracellular matrix accumulation compared to non-degradable hydrogels (collagen type II; 3.2-fold increase, aggrecan; 4-fold increase, SOX9; 2.8-fold increase). Hydrogels containing a low concentration of the RGDS peptide displayed significantly decreased collagen type I and X gene expression profiles, suggesting a major advantage over either hydrogels functionalized with a higher RGDS peptide concentration, or non-degradable hydrogels, in promoting an articular cartilage phenotype. These highly versatile Scl2 hydrogels can be further manipulated to improve specific elements of the chondrogenic response by hMSCs, through the introduction of additional bioactive and/or biodegradable motifs. As such, these hydrogels have the possibility to be used for other applications in tissue engineering.

Recapitulating aspects of the native tissue biochemical microenvironment faces significant challenges in regenerative medicine and tissue engineering due to the complex and dynamic nature of the tissue. The ability to take advantage of, mimic, and modulate cell-mediated processes within novel naturally-derived hydrogels is of great interest in the field of biomaterials to generate constructs that more closely resemble the biochemical microenvironment and functions of native biological tissues such as articular cartilage. Towards this goal, the temporal presentation of bioactive sequences such as RGDS on the chondrogenic differentiation of human mesenchymal stem cells is considered important as it has been shown to influence the chondrogenic phenotype. Here, a novel and versatile platform to recreate a high degree of biological complexity is proposed, which could also be applicable to other tissue engineering and regenerative medicine applications.

Partial Text

Articular cartilage is a highly complex connective tissue that covers the surface of bones in synovial joints [1]. The unique spatial organization of the components of cartilage extracellular matrix is fundamental to its ability to carry out its biomechanical functions [2], [3]. Trauma to articular cartilage and/or disease of the joint can stimulate catabolic responses that disturb tissue homeostasis and can lead to progressive degeneration [4]. This is aggravated by the aneural and avascular nature of articular cartilage, combined with the limited ability of resident cells to migrate to sites of injury, which contribute to a restricted capacity for self-repair and regeneration [5]. Current clinical treatments for articular cartilage ailments, such as non-steroidal anti-inflammatory drugs [6], viscosupplementation [4], mosaicplasty [7], autologous chondrocyte implantation [8], microfracture [9], and periosteal transplantation [10], generally provide short-term pain relief and recovery of joint mobility to patients, but long-term benefits often remain elusive [3]. The repair tissue formed as a result of the surgical interventions listed here often does not exhibit the same biochemical composition as native tissue, leading to inferior biomechanical properties. Repair tissue is typically rapidly degenerated, ultimately leading to the failure of the intervention [4], thus requiring additional treatment and eventually total joint arthroplasty [11].

In this work, we have developed novel hydrogels based on collagen-mimetic proteins with cell-mediated control over the temporal presentation of bioactive cues, in this case the RGDS peptide. The enzymatic release of tethered RGDS from a hydrogel that also incorporated peptide sequences to dynamically, specifically, and non-covalently bind heparin, HA, and CS was shown to significantly promote hMSC chondrogenesis compared to hydrogels for which the RGDS peptide remained tethered to the backbone throughout the culture period. Additionally, hydrogels functionalized with a low concentration of the RGDS peptide resulted in decreased collagen type I and X expression. The results in this work highlight the importance of the temporal presentation of bioactive cues and present an approach to achieve this degree of complexity. This work presents an additional example of the architectural flexibility and modularity of our novel collagen-mimetic hydrogel platform and its potential utility in tissue engineering and regenerative medicine applications.




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