Decellularized sturgeon cartilage extracellular matrix scaffold inhibits chondrocyte hypertrophy in vitro and in vivo

Abstract

Since chondrocyte hypertrophy greatly limits the efficiency of cartilage defects repairing via cartilage tissue engineering (CTE), it is critical to develop a functional CTE scaffold able to inhibit chondrocyte hypertrophy during this period of cartilage regeneration. In this study, we tested the applicability of using decellularized sturgeon cartilage ECM (dSCECM) scaffold to cease chondrocyte hypertrophy during cartilage damage repair. The dSCECM scaffolds with interconnected porous structure and pore size of 114.1 ± 20.9 μm were successfully prepared with freeze‐dry method. Chondrocytes displayed a round shape and aggregated to form cellular spheroids within dSCECM scaffolds, which is similar to their chondrocytic phenotype within cartilage in vivo. Higher transcriptional level of chondrogenic related genes and integrin related genes was observed in chondrocytes incubated with dSCECM scaffolds instead of type I collagen (COL I) scaffolds, which were used as the control due to their widely usage in CTE and clinic applications. Furthermore, it confirmed that, compared with COL I scaffolds, dSCECM scaffolds significantly reduced the transcription of chondrocyte hypertrophy related genes in chondrocytes following the hypertrophic induction treatment. To test the ability of dSCECM scaffold to inhibit chondrocytes hypertrophy in vivo, chondrocytes with dSCECM scaffolds and COL I scaffolds were cultured with hypertrophic media and were implanted into nude mice respectively. Following 4 weeks implantation, interestingly, only the specimens derived from COL I scaffolds displayed consequences of chondrocyte hypertrophy like calcification deposition, demonstrating that chondrocyte hypertrophy is ceased by the dSCECM scaffold following hypertrophic induction. It suggests that the dSCECM scaffold can be potentially applied in clinical treating cartilage defects via the CTE approach to avoid the risk of chondrocyte hypertrophy.

Cartilage Repair Using Clematis Triterpenoid Saponin Delivery Microcarrier, Cultured in a Microgravity Bioreactor Prior to Application in Rabbit Model

“…Chondrocytes are one of the classic seed cell types in CTE, but they are prone to dedifferentiation in the process of in vitro amplification. Many scholars have found that technological improvements such as addition of growth factors and 3D biological scaffolds can effectively reverse chondrocyte dedifferentiation. , In the process of constructing tissue-engineered cartilage, the ideal biological scaffold should not only provide a 3D structural environment for chondrocyte adhesion, growth, and interaction but also create a cellular biological microenvironment conducive to cartilage repair as much as possible and provide appropriate growth factors to maintain the activity of chondrocytes, which is the key to cartilage regeneration. , …”

Generation and Evaluation of Novel Biomaterials Based on Decellularized Sturgeon Cartilage for Use in Tissue Engineering

“…In the present work we used sturgeon cartilage as a biomaterial in TE. Although this biomaterial has been very recently used in cartilage TE, although using a different approach [21,22], its biological use is still preliminary and further research is in need. In general, this material offers several advantages compared to other types of scaffold.…”

“…In general, the use of xenografts obtained from other species requires previous decellularization in order to eliminate all non-human cells without significantly altering the ECM [28,37,49]. In the present work, we applied a combination of several previously described physical, chemical and enzymatic decellularization methods [27][28][29] to sturgeon cartilage, as recently described for different species of sturgeon [21,22]. As a highly dense, compact tissue, cartilage is considered to be difficult to decellularize, mostly because of the low penetrability of decellularization agents in the dense ECM, and thus their limited efficiency in removing all cells and cell debris [12].…”

“…Previous work has identified some of the molecules in sturgeon ECM, including collagen [18,19] and chondroitin sulfate [20], which have been isolated and characterized. Although very little information is available on the general structure and composition of sturgeon cartilage, this material has been recently used in TE with promising results [21,22].…”

Preparation and Use of Decellularized Extracellular Matrix for Tissue Engineering

“…Fibroblasts have been shown to be more responsive to their microenvironment when grown in skin dECM bioink compared to type I collagen [ 107 ]. Cartilage dECM has been shown to be effective at preventing chondrocyte hypertrophy and calcification of the cartilage in the repair of cartilage defects [ 108 ]. The ability to improve cell viability and function and promote cell proliferation has also been shown in other cell types when cultured on dECM and, especially, tissue-specific dECM [ 14 , 56 , 82 , 93 , 98 , 109 , 110 , 111 , 112 , 113 , 114 , 115 ].…”