Regulation of Extracellular Matrix Synthesis by Shell Extracts from the Marine Bivalve Pecten maximus in Human Articular Chondrocytes— Application for Cartilage Engineering
The shells of the bivalve mollusks are organo-mineral structures predominantly composed of calcium carbonate, but also of a minor organic matrix, a mixture of proteins, glycoproteins, and polysaccharides. These proteins are involved in mineral deposition and, more generally, in the spatial organization of the shell crystallites in well-defined microstructures. In this work, we extracted different organic shell extracts (acid-soluble matrix, acid-insoluble matrix, water-soluble matrix, guanidine HCl/EDTA-extracted matrix, referred as ASM, AIM, WSM, and EDTAM, respectively) from the shell of the scallop Pecten maximus and studied their biological activities on human articular chondrocytes (HACs). We found that these extracts differentially modulate the biological activities of HACs, depending on the type of extraction and the concentration used. Furthermore, we showed that, unlike ASM and AIM, WSM promotes maintenance of the chondrocyte phenotype in monolayer culture. WSM increased the expression of chondrocyte-specific markers (aggrecan and type II collagen), without enhancing that of the main chondrocyte dedifferentiation marker (type I collagen). We also demonstrated that WSM could favor redifferentiation of chondrocyte in collagen sponge scaffold in hypoxia. Thus, this study suggests that the organic matrix of Pecten maximus, particularly WSM, may contain interesting molecules with chondrogenic effects. Our research emphasizes the potential use of WSM of Pecten maximus for cell therapy of cartilage.
KeywordsPecten maximus Shell extracts Water-soluble matrix Human articular chondrocytes Cartilage engineering
The project received the label of AQUIMER pole (M. Coquelle). The authors thank Copalis (Boulogne-Sur-Mer, France) for providing the shell powders.
MB, JML, PG, and FL designed the experiments. MB, RR, TGL, TL, and FL performed the experiments. MB, AS, JML, PG, and FL analyzed the data. FC, FM, and SL contributed the reagents, materials, and analysis tools. MB and FL wrote the paper. AS, FM, and PG realized the critical revision of the article. JML and PG provided the financial support.
This work was financially supported by the “Fonds Unique Interministériel” (FUI, French Government) through the SEAMINEROIL program to PG and JML. The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. MB: Fellowship from the Regional Council of Lower Normandy. TGL: Fellowship from the French Ministry of Research and Technology. TL: Fellowship supported by the SEAMINEROIL grant from the FUI. FL: Post-doctoral fellow supported by a 1-year grant from the FUI.
Compliance with Ethical Standards
All patients signed an informed consent agreement form, which was approved by the local ethics committee.
Conflict of Interest
The authors declare that they have no conflicts of interest.
- Almeida MJ, Milet C, Peduzzi J, Pereira L, Haigle J, Barthelemy M, Lopez E (2000) Effect of water-soluble matrix fraction extracted from the nacre of Pinctada maxima on the alkaline phosphatase activity of cultured fibroblasts. J Exp Zool 288:327–334Google Scholar
- Demoor M, Maneix L, Ollitrault D, Legendre F, Duval E, Claus S, Mallein-Gerin F, Moslemi S, Boumediene K, Galera P (2012) Deciphering chondrocyte behaviour in matrix-induced autologous chondrocyte implantation to undergo accurate cartilage repair with hyaline matrix. Pathol Biol 60:199–207CrossRefPubMedGoogle Scholar
- Demoor M, Ollitrault D, Gomez-Leduc T, Bouyoucef M, Hervieu M, Fabre H, Lafont J, Denoix JM, Audigié F, Mallein-Gerin F, Legendre F, Galera P (2014) Cartilage tissue engineering: molecular control of chondrocyte differentiation for proper cartilage matrix reconstruction. Biochim Biophys Acta 1840:2414–2440CrossRefPubMedGoogle Scholar
- Latire T, Legendre F, Bigot N, Carduner L, Kellouche S, Bouyoucef M, Carreiras F, Marin F, Lebel JM, Galéra P, Serpentini A (2014) Shell extracts from the marine bivalve Pecten maximus regulate the synthesis of extracellular matrix in primary cultured human skin fibroblasts. PLoS One 9:e99931CrossRefPubMedPubMedCentralGoogle Scholar
- Legendre F, Heuze A, Boukerrouche K, Leclercq S, Boumediene K, Galera P, Domagala F, Pujol JP, Ficheux H (2009) Rhein, the metabolite of diacerhein, reduces the proliferation of osteoarthritic chondrocytes and synoviocytes without inducing apoptosis. Scand J Rheumatol 38:104–111CrossRefPubMedGoogle Scholar
- Legendre F, Ollitrault D, Hervieu M, Baugé C, Maneix L, Goux D, Chajra H, Mallein-Gerin F, Boumediene K, Galera P, Demoor M (2013) Enhanced hyaline cartilage matrix synthesis in collagen sponge scaffolds by using siRNA to stabilize chondrocytes phenotype cultured with bone morphogenetic protein-2 under hypoxia. Tissue Eng Part C Methods 19:550–567CrossRefPubMedGoogle Scholar
- Mouriès LP, Almeida M-J, Milet C et al (2002) Bioactivity of nacre water-soluble organic matrix from the bivalve mollusk Pinctada maxima in three mammalian cell types: fibroblasts, bone marrow stromal cells and osteoblasts. Comp Biochem Physiol B Biochem Mol Biol 132:217–229CrossRefPubMedGoogle Scholar
- Ollitrault D, Legendre F, Drougard C, Briand M, Benateau H, Goux D, Chajra H, Poulain L, Hartmann D, Vivien D, Shridhar V, Baldi A, Mallein-Gerin F, Boumediene K, Demoor M, Galera P (2015) BMP-2, hypoxia, and COL1A1/HtrA1 siRNAs favor neo-cartilage hyaline matrix formation in chondrocytes. Tissue Eng Part C Methods 21:133–147CrossRefPubMedGoogle Scholar
- Torita A, Miyamoto A, Hasegawa Y (2007) The effects of scallop shell extract on collagen synthesis. Fish Sci 73:1388–1394Google Scholar