Osteogenesis of Human iPSC-Derived MSCs by PLLA/SF Nanofiber Scaffolds Loaded with Extracellular Matrix

Bone defects that arise from trauma, skeletal diseases, or tumor resections have become the commonest and most thorny problems in orthopedic clinics. Recently, biocomposite materials used as artificial bone repair materials have provided a promising approach for bone regeneration. In this study, poly (l-lactide acid) (PLLA) and silk fibroin (SF) were used to fabricate nanofiber scaffolds by electrospinning technology. In order to simulate a biomimetic osteoblast microenvironment, decellularized extracellular matrix from osteoblasts was loaded into the biocomposite scaffolds (O-ECM/PLLA/SF). It was found that the O-ECM/PLLA/SF scaffolds were nontoxic for L929 cells and had good cytocompatibility. Their effects on mesenchymal stem cells derived from human-induced pluripotent stem cell (iPSC-MSC) behavior were investigated. As a result, the scaffolds with the addition of O-ECM showed enhanced alizarin red S (ARS) activity. In addition, higher expression of osteogenic gene markers such as runt-related transcription factor 2 (Runx2), collagen type I (Col-1), and osteocalcin (OCN) as well as upregulated expression of osteogenic marker protein osteopontin (OPN) and Col-1 further substantiated the applicability of O-ECM/PLLA/SF scaffolds for osteogenesis. Furthermore, the in vivo study also indicated maximal new bone formation in the skull defect model of Sprague Dawley (SD) rats treated with the O-ECM/PLLA/SF carried by human iPSC-MSCs. Hence, this study suggests that O-ECM/PLLA/SF scaffolds have a potential application in bone tissue engineering.

[1]  Jun Luo,et al.  Alginate-modified mesoporous bioactive glass and its drug delivery, bioactivity, and osteogenic properties , 2022, Frontiers in Bioengineering and Biotechnology.

[2]  S. Dong,et al.  Staggered Nanofiber Scaffolds via Electric-Field-Controlled Assembly for Bone Tissue Regeneration , 2022, ACS Applied Nano Materials.

[3]  Shaohua Wu,et al.  Electrospun strong, bioactive, and bioabsorbable silk fibroin/poly (L-lactic-acid) nanoyarns for constructing advanced nanotextile tissue scaffolds , 2022, Materials today. Bio.

[4]  X. Lou,et al.  Osteoblast-derived extracellular matrix coated PLLA/silk fibroin composite nanofibers promote osteogenic differentiation of bone mesenchymal stem cells. , 2021, Journal of biomedical materials research. Part A.

[5]  M. Iqbal,et al.  Fabrication of robust poly l-lactic acid/cyclic olefinic copolymer (PLLA/COC) blends: study of physical properties, structure, and cytocompatibility for bone tissue engineering , 2021, Journal of Materials Research and Technology.

[6]  Zihan Lu,et al.  Fabrication of hierarchical porous poly (l-lactide) (PLLA) fibrous membrane by electrospinning , 2021 .

[7]  Mengguang Fu,et al.  Design and research of bone repair scaffold based on two-way fluid-structure interaction , 2021, Comput. Methods Programs Biomed..

[8]  Zhihong Wang,et al.  Biofabrication of poly(l-lactide-co-ε-caprolactone)/silk fibroin scaffold for the application as superb anti-calcification tissue engineered prosthetic valve. , 2021, Materials science & engineering. C, Materials for biological applications.

[9]  Hongbing Deng,et al.  Carboxymethyl chitosan/sodium alginate-based micron-fibers fabricated by emulsion electrospinning for periosteal tissue engineering , 2020 .

[10]  Huawei Qu.  Additive manufacturing for bone tissue engineering scaffolds , 2020 .

[11]  A. Padalhin,et al.  Functionalization of extracellular matrix (ECM) on multichannel biphasic calcium phosphate (BCP) granules for improved bone regeneration , 2020 .

[12]  Kwideok Park,et al.  Novel ECM Patch Combines Poly(vinyl alcohol), Human Fibroblast-Derived Matrix, and Mesenchymal Stem Cells for Advanced Wound Healing. , 2020, ACS biomaterials science & engineering.

[13]  Gerry L. Koons,et al.  Materials design for bone-tissue engineering , 2020, Nature Reviews Materials.

[14]  Y. Wang,et al.  Stereocomplex poly(lactic acid)-based composite nanofiber membranes with highly dispersed hydroxyapatite for potential bone tissue engineering , 2020 .

[15]  Renze Shen,et al.  Emulsion electrospun PLA/calcium alginate nanofibers for periodontal tissue engineering , 2020, Journal of biomaterials applications.

[16]  F. Wang,et al.  Silk fibroin-poly(lactic acid) biocomposites: Effect of protein-synthetic polymer interactions and miscibility on material properties and biological responses. , 2019, Materials science & engineering. C, Materials for biological applications.

[17]  Yueping Chen,et al.  Biocompatiable silk fibroin/carboxymethyl chitosan/strontium substituted hydroxyapatite/cellulose nanocrystal composite scaffolds for bone tissue engineering. , 2019, International journal of biological macromolecules.

[18]  Anuj Kumar,et al.  Development of biomimetic electrospun polymeric biomaterials for bone tissue engineering. A review , 2019, Journal of biomaterials science. Polymer edition.

[19]  D. Vashishth,et al.  Co-culture cell-derived extracellular matrix loaded electrospun microfibrous scaffolds for bone tissue engineering. , 2019, Materials science & engineering. C, Materials for biological applications.

[20]  Hongwei Lu,et al.  Functionalized cell-free scaffolds for bone defect repair inspired by self-healing of bone fractures: A review and new perspectives. , 2019, Materials Science and Engineering C: Materials for Biological Applications.

[21]  Younan Xia,et al.  Electrospinning and Electrospun Nanofibers: Methods, Materials, and Applications. , 2019, Chemical reviews.

[22]  Xiaofeng Jia,et al.  Three-dimensional (3D) printed scaffold and material selection for bone repair. , 2019, Acta biomaterialia.

[23]  X. Mo,et al.  Three-dimensional electrospun nanofibrous scaffolds displaying bone morphogenetic protein-2-derived peptides for the promotion of osteogenic differentiation of stem cells and bone regeneration. , 2019, Journal of colloid and interface science.

[24]  Huiming Wang,et al.  Cell-Sheet-Derived ECM Coatings and Their Effects on BMSCs Responses. , 2018, ACS applied materials & interfaces.

[25]  T. Tian,et al.  Electrospun Poly(3-hydroxybutyrate-co-4-hydroxybutyrate)/Graphene Oxide Scaffold: Enhanced Properties and Promoted in Vivo Bone Repair in Rats. , 2017, ACS applied materials & interfaces.

[26]  Mark A. Lee,et al.  Extracellular Matrix-Coated Composite Scaffolds Promote Mesenchymal Stem Cell Persistence and Osteogenesis. , 2016, Biomacromolecules.

[27]  B. Han,et al.  Composite poly(l-lactic-acid)/silk fibroin scaffold prepared by electrospinning promotes chondrogenesis for cartilage tissue engineering , 2016, Journal of biomaterials applications.

[28]  Richard O.C. Oreffo,et al.  Biofabrication of bone tissue: approaches, challenges and translation for bone regeneration. , 2016, Biomaterials.

[29]  Keita Ito,et al.  Silk fibroin as biomaterial for bone tissue engineering. , 2016, Acta biomaterialia.

[30]  Yanzhong Zhang,et al.  Well-aligned chitosan-based ultrafine fibers committed teno-lineage differentiation of human induced pluripotent stem cells for Achilles tendon regeneration. , 2015, Biomaterials.

[31]  H. Yatani,et al.  Comparative analysis of mouse-induced pluripotent stem cells and mesenchymal stem cells during osteogenic differentiation in vitro. , 2014, Stem cells and development.

[32]  Huijing Zhao,et al.  Preparation of uniaxial multichannel silk fibroin scaffolds for guiding primary neurons. , 2012, Acta biomaterialia.

[33]  F. Locatelli,et al.  Mesenchymal stromal cell therapy: a revolution in Regenerative Medicine? , 2012, Bone Marrow Transplantation.

[34]  B. Gramsch,et al.  Growth and Differentiation of Osteoblast‐Like Cells from Calvaria of Connexin43 Deficient Mice , 2004 .

[35]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[36]  V. Chiono,et al.  Intermolecular interactions between B. mori silk fibroin and poly(l-lactic acid) in electrospun composite nanofibrous scaffolds. , 2017, Materials science & engineering. C, Materials for biological applications.

[37]  Yanzhong Zhang,et al.  Osteogenic differentiation and bone regeneration of iPSC-MSCs supported by a biomimetic nanofibrous scaffold. , 2016, Acta biomaterialia.

[38]  R L Juliano,et al.  Signal transduction by cell adhesion receptors and the cytoskeleton: functions of integrins, cadherins, selectins, and immunoglobulin-superfamily members. , 2002, Annual review of pharmacology and toxicology.