Low-Molecular-Weight Heparin-Functionalized Chitosan-Chondroitin Sulfate Hydrogels for Controlled Release of TGF-β3 and in vitro Neocartilage Formation

Abstract

Repair of hyaline cartilage remains a huge challenge in clinic because of the avascular and aneural characteristics and the paucity of endogenous repair cells. Recently, tissue engineering technique, possessing unique capacity of repairing large tissue defects, avoiding donor complications and two-stage invasive surgical procedures, has been developed a promising therapeutic strategy for cartilage injury. In this study, we incorporated low-molecular-weight heparin (LMWH) into carboxymethyl chitosan-oxidized chondroitin sulfate (CMC-OCS) hydrogel for loading transforming growth factor-β3 (TGF-β3) as matrix of peripheral blood mesenchymal stem cells (PB-MSCs) to construct tissue-engineered cartilage. Meanwhile, three control hydrogels with or without LMWH and/or TGF-β3 were also prepared. The gelling time, microstructures, mechanical properties, degradation rate, cytotoxicity, and the release of TGF-β3 of different hydrogels were investigated. In vitro experiments evaluated the tri-lineage differentiation potential of PB-MSCs, combined with the proliferation, distribution, viability, morphology, and chondrogenic differentiation. Compared with non-LMWH-hydrogels, LMWH-hydrogels (LMWH-CMC-OCS-TGF-β3) have shorter gelling time, higher mechanical strength, slower degradation rate and more stable and lasting release of TGF-β3. After two weeks of culture in vitro, expression of cartilage-specific genes collagen type-2 (COL-2) and aggrecan (AGC), and secretion of glycosaminoglycan (GAG), and COL-2 proteins in LMWH-CMC-OCS-TGF-β3 group were significantly higher than those in other groups. COL-2 immunofluorescence staining showed that the proportion of COL-2 positive cells and immunofluorescence intensity in LMWH-CMC-OCS-TGF-β3 hydrogel were significantly higher than those in other groups. The LMWH-CMC-OCS-TGF-β3 hydrogel can slowly release TGF-β3 in a long term, and meanwhile the hydrogel can provide a biocompatible microenvironment for the growth and chondrogenic differentiation of PB-MSCs. Thus, LMWH functionalized CMC-OCS hydrogels proposed in this work will be beneficial for constructing functional scaffolds for tissue-engineered cartilage.

The Use of Peripheral Blood-Derived Stem Cells for Cartilage Repair and Regeneration In Vivo: A Review

“…For example, MSCs derived from bone marrow, synovium or adipose tissue reached 80-90% confluence within 7 to 14 days (Zhang et al, 2014;Jin et al, 2016;Shimomura et al, 2016). However, MSCs derived from PB did not achieve the same confluence until about 21 days after primary culture (Chen et al, 2019). It takes longer to obtain the culture-expanded PB-MSCs than other tissue-derived MSCs.…”

“…Compared with other tissue-derived MSCs, the culture of PB-MSCs was relatively difficult, which resulted in less reports of its application in vivo, but it does not affect its application prospects. On the contrary, it is ethically more suitable for clinical application due to its unique advantages, such as minimally invasive sample acquisition procedure, repeatable sampling, and high recognition of patients (Fu et al, 2014a;Fu et al, 2014b;Wang et al, 2016a;Chen et al, 2019). In this review, we have summarized all the currently published researches on the use of PBSCs for cartilage repair and regeneration in vivo.…”

Progressive 3D Printing Technology and Its Application in Medical Materials

“…Multiple bioprinting applications in vascular models, soft-tissue models, and bone models manufactured with extrusion-based printing technology have been well-developed in recent years (Ahlfeld et al, 2017;Paxton et al, 2017;Ahlfeld et al, 2018). One major advantage of its bioprinting application is that the hydrogels of extrusion-based printing is capable to fabricate products with high cell density (> 1 × 10 6 cells ml −1 ) (Petta et al, 2018;Taylor et al, 2018;Chen et al, 2019).…”

Advancements and Frontiers in the High Performance of Natural Hydrogels for Cartilage Tissue Engineering

“…Chitosan is a kind of mucopolysaccharide widely existed in nature, which is important component of connective tissue with complexation, bacteriostasis, adsorption, and antioxidant effects (Molinaro et al, 2002;Jayakumar et al, 2005). Recent reports demonstrated that chitosan can be gelated in an acidic pH or a non-solvent condition (Ribeiro et al, 2017;Xu et al, 2017;Chen Y. R. et al, 2019), and further be prepared for chitosanbased hydrogel scaffolds. Chitosan has good biocompatibility and biodegradability; therefore, it is a kind of tissue-engineering material with wide application prospects and can be considered as a potential material for cartilage repair in regenerative medicine fields.…”