Evaluation of Chondrogenic Differentiation Ability of Bone Marrow Mesenchymal Stem Cells in Silk Fibroin/Gellan Gum Hydrogels Using miR-30

Eun Yeong Shin; Jong Ho Park; Myeong Eun Shin; Jeong Eun Song; Cristiano Carlomagno; Gilson Khang

Macromolecular Research, Vol.27, No.4, 369-376, April, 2019

DOI10.1007/s13233-019-7048-x Export Citation

Evaluation of Chondrogenic Differentiation Ability of Bone Marrow Mesenchymal Stem Cells in Silk Fibroin/Gellan Gum Hydrogels Using miR-30

Eun Yeong Shin, Jong Ho Park, Myeong Eun Shin, Jeong Eun Song, Cristiano Carlomagno¹, and Gilson Khang^†

Department of BIN Convergence Technology, Department of Polymer Nano Science & Technology and Polymer Materials Fusion Research Center, Chonbuk National University, 567 Baekje-daero, Deokjin-gu, Jeonju-si, Jeonbuk 54896, Korea
¹Department of Industrial Engineering, University of Trento, via Sommarive 9, Trento, Italy BIOTech

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The poor proliferative ability of chondrocytes makes complicated the cartilage regeneration after injuries or during the pathological state. Nowadays, stem cells represent a potential tool for different tissues regeneration, including cartilage. Previous studies demonstrated the role of miRNAs (MicroRNAs) in chondrogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) through the in inhibition of specific genes expression. In this study, miR-30a was used to assess the possible chondrogenic differentiation, in combination with silk fibroin/gellan gum (SF/GG) hydrogels, suitable for cells sustaining and proliferation. The SF/GG hydrogel was fabricated combining 2% of gellan gum with 2% of silk fibroin, exploiting the cationic cross-linking of the polysaccharide. For characterization, the hydrogel was lyophilized and used. Scanning electron microscopy was used to characterize the scaffold morphology, while FT-IR spectroscopy was performed to evaluate the chemical properties. Suitability of the produced scaffold for cells adhesion and nutrient and oxygen perfusion was evaluated through water uptake and overall porosity. BMSCs extracted from rats were transfected with miR-30a mimic and inhibitor. MiR-30a expression rates were measured by real time-quantitative polymerase chain reaction (qPCR) monitoring the expression of cartilage-specific gene through reverse transcription-polymerase chain reaction (RT-PCR). Histological assays were used to identify the chondrogenesis of BMSCs on the SF/GG hydrogel. Our results demonstrated the suitability of the SF/GG hydrolgel for cells adhesion, ingrowth and nutrients perfusion. The exposition of cells to the miR-30a demonstrated the potential role of the molecule in chondrogenic differentiation showing an up regulation of cartilage-specific gene. In conclusion, stem cells transfected with miRNA can positively affect articular cartilage regeneration and the potential of BMSCs-encapsulated hydrogel transfected with miR-30a as a therapeutics for osteoarthritis (OA) has been confirmed.

Keywords:miR-30a;chondrogenesis;SF/GG hydrogel;articular cartilage;tissue engineering

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[2] Marlovits S, Zeller P, Singer P, Resinger C, Vecsei V, Eur. J. Radiol., 57, 24 (2006)

[3] Wakitani S, Goto T, Pineda SJ, Young RG, Mansour JM, Caplan AI, Goldberg VM, J. Bone Joint Surg. Am., 76, 579 (1994)

[4] Williams CG, Kim TK, Taboas A, Malik A, Manson P, Elisseeff J, Tissue Eng., 9, 679 (2003)

[5] Johnstone B, Hering TM, Caplan AI, Goldberg VM, Yoo JU, Exp. Cell Res., 238, 265 (1998)

[6] Murdoch AD, Grady LM, Ablett MP, Katopodi T, Meadows RS, Hardingham TE, Stem. Cells., 25, 2786 (2007)

[7] Oliveira JT, Gardel LS, Martins L, Gomes ME, Reis RL, J. Orthop. Res., 28, 1193 (2010)

[8] Oliveira JT, Santos TC, Martins L, Picciochi R, Marques AP, Castro AG, Neves NM, Mano, Reis RL, Tissue Eng. Part A, 16, 343 (2010)

[9] Wang Y, Kim HJ, Vunjak-Novakovic G, Kaplan DL, Biomaterials, 27, 6064 (2006)

[10] Koh LD, Cheng Y, Teng CP, Khin YW, Loh XJ, Tee SY, Low M, Ye E, Yu HD, Zhang YW, Han MY, Prog. Polym. Sci, 46, 86 (2015)

[11] Rockwood DN, Preda RC, Yucel T, Wang X, Lovett ML, Kaplan DL, Nat. Protoc., 6, 1612 (2011)

[12] Park JH, Jeon HY, Jeon YS, Park H, Kim CM, Song JE, Khang GS, Polym. Korea, 42(2), 298 (2018)

[13] Chen Y, Stallings RL, Cancer Res., 67, 976 (2007)

[14] Mao G, Zhang Z, Huang Z, Chen W, Huang G, Meng F, Zhang Z, Kang Y, Osteoarthr. Cartil., 25, 521 (2017)

[15] Paik S, Jung HS, Lee S, Yoon DS, Park MS, Lee JW, Stem Cells Dev., 21, 3298 (2012)

[16] Zhou X, Wang J, Sun H, Qi Y, Xu W, Luo D, Jin X, Li C, Chen W, Lin Z, Li F, Zhang R, Li G, Cell Tissue Res., 366, 143 (2016)

[17] Xue ZL, Meng YL, Ge JH, Exp. Ther. Med., 14, 1481 (2017)

[18] Tian Y, Guo R, Shi B, Chen L, Yang L, Fu Q, Life Sci., 148, 220 (2016)

[19] De Laporte L, Shea LD, Adv. Drug Deliv. Rev., 59, 292 (2007)

[20] Loh QL, Choong C, Tissue Eng. Part B: Rev., 19, 485 (2013)

[21] Kim DK, Kim JI, Hwang TI, Sim BR, Khang G, ACS Appl. Mater. Interfaces, 9, 1384 (2017)

[22] Anseth KS, Bowman CN, BrannonPeppas L, Biomaterials, 17, 1647 (1996)

[23] Johnson BD, Beebe DJ, Crone W, Mater. Sci. Eng. C-Biomimetic Supramol. Syst., 24, 575 (2004)

[24] Sung HJ, Meredith C, Johnson C, Galis ZS, Biomaterials, 25, 5735 (2004)

[25] Lu QA, Zhang B, Li MZ, Zuo BQ, Kapan DL, Huang YL, Zhu HS, Biomacromolecules, 12(4), 1080 (2011)

[26] Lee KS, Kim HJ, Li QL, Chi XZ, Ueta C, Komori T, Wozney JM, Kim EG, Choi JY, Ryoo HM, Bae SC, Mol. Cell. Biochem., 20, 8783 (2000)

[27] Bi WM, Deng JM, Zhang ZP, Behringer RR, de Crombrugghe B, Nat. Genet., 22, 85 (1999)

[28] Watanabe H, Yamada Y, Kimata K, J. Biochem-Tokyo., 124, 687 (1998)