Article
  • VEGF/Laminin Co-Immobilized Membrane for Enhanced Differentiation of Muscle-derived Stem Cells into Smooth Muscle Cells

  • So Ri Lee, Tae Ho Kim, Se Heang Oh*,**, Seong Keun Kwon***, and Jin Ho Lee†

  • Department of Advanced Materials and Chemical Engineering, Hannam University, Daejeon 34054, Korea
    *Department of Nanobiomedical Science, Dankook University, Cheonan 31116, Korea
    **Department of Pharmaceutical Engineering, Dankook University, Cheonan 31116, Korea
    ***Department of Otorhinolaryngology-Head and Neck Surgery, Seoul National University Hospital, Seoul 03080, Korea

  • 근육유래 줄기세포의 평활근으로 분화유도를 위한 VEGF/Laminin 탑재 맴브레인

  • 이소리 · 김태호 · 오세행*,** · 권성근*** · 이진호†

  • 한남대학교 화공신소재공학과, *단국대학교 나노바이오의과학과, **단국대학교 제약공학과, ***서울대학교병원 이비인후과

Abstract

It is important to control the stem cell differentiation into target cell types for clinical use of the stem cells. In this study, asymmetrically porous polycaprolactone (PCL)/Pluronic F127 membranes immobilized with bioactive molecules [vascular endothelial growth factor (VEGF) and/or laminin] were prepared to investigate the effect of musclederived stem cells (MDSCs) on smooth muscle cell (SMC) differentiation. The VEGF and laminin immobilized on the PCL/F127 membrane surface were released with a sustained manner for 28 and 14 days from the membrane, respectively. The SMC differentiation behavior of MDSCs on the membranes immobilized with single or dual bioactive molecules was compared by DNA quantification, RT-PCR, and immunohistochemical analyses. The dual VEGF/laminin-immobilized membrane group showed higher cell growth and more effective SMC differentiation than the single VEGF- or lamininimmobilized group. From our findings, we suggest that the dual VEGF/laminin-immobilized membrane may be applicable to use as a guided smooth muscle regeneration membrane.


본 연구에서는 생리활성인자[vascular endothelial growth factor(VEGF) 및 laminin]가 탑재된, 비대칭구조를 가지는 다공성 polycaporlactone(PCL) 기반의 맴브레인을 제조하였으며, 맴브레인에 탑재된 생리활성인자에 의한 근육유래 줄기세포(muscle-derived stem cells, MDSCs)의 평활근세포(smooth muscle cells, SMCs)로의 분화거동을 연구하였다. PCL 기반 맴브레인에 탑재된 VEGF 및 laminin은 맴브레인으로부터 각각 28일과 14일 동안 지속적으로 방출되었다. 각 생리활성인자가 단독 혹은 공동으로 탑재된 PCL 기반 맴브레인에서 MDSCs의 SMCs로의 분화거동을 DNA 정량화, RT-PCR 및 면역염색을 통해 분석하였으며, 두 생리활성인자가 공동으로 탑재된 PCL 기반 맴브레인에서 가장 우수한 평활근으로의 분화거동을 보임을 확인하였다. 이러한 결과로부터, VEGF/laminin이 탑재된, 비대칭구조를 가지는 다공성 PCL 기반의 맴브레인은 평활근 재생을 위한 재생막으로 사용이 가능할 것이라 판단되었다.


Keywords: porous membrane, vascular endothelial growth factor, laminin, stem cell, differentiation

Introduction

Stem cells are widely used in tissue engineering for the regeneration of various tissues or organs.1,2 It is well known that the stem cells are differentiated into various cell types, such as bone marrow stem cells into osteogenic, chondrogenic, myogenic, vascular, and neurogenic cells;3,4 adipose stem cells into osteogenic, myogenic, and chondrogenic cells;5,6 skin stem cells into neurons and smooth muscle cells;6 and musclederived stem cells (MDSCs) into smooth muscle cells (SMCs).7-9 Various growth factors and cytokines are commonly adapted to achieve the differentiation of stem cells into specific target cells. It is important to precisely control the stem cell differentiation into specific cell types for the clinical use of stem cells. Particularly, the MDSCs capable of differentiating into SMCs have been widely used for the smooth muscle tissue regeneration in urethra,10 anus,11,12 and bladder.13 In our previous studies,14,15 dual growth factors [vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF)]-immobilized polycaprolactone (PCL)/Pluronic F127 beads were prepared as an injectable bioactive urethral bulking agent. Both bFGF and VEGF are well known as the growth factors which induce smooth muscle differentiation9,16 as well as angiogenesis. It was observed that the growth factors (bFGF and VEGF)-immobilized porous beads stimulate SMC differentiation of MDSCs and thus defect tissue regeneration around urethra (mainly smooth muscle) to improve the sphincter function.
In this study, we prepared dual bioactive molecules [VEGF (growth factor) and laminin (protein)]-immobilized PCL/Pluronic F127 asymmetrically porous membrane to use as a guided tissue (smooth muscle) regeneration membrane. Laminin is the major protein of the basal lamina of various tissues, and is known to be a key protein responsible for cell migration and differentiation.17,18 In particular, it is known that the laminin plays important roles in SMC differentiation and maintenance of the SMC phenotype.19 We tested whether using cell differentiation-stimulating growth factor (VEGF) and protein (laminin) together provides a synergistic effect on SMC differentiation of MDSCs on the membrane surface. VEGF, which is one of heparin-binding growth factor families, was loaded on the surface of porous PCL/Pluronic F127 membrane via specific interactions between heparin and Pluronic F127 (hydrogen bonding) and subsequent interactions between growth factor and heparin (ionic interaction), which preserve the biological activity of growth factor without denaturation. Laminin was immobilized on the membrane pore surface by physical adsorption. The VEGF and laminin immobilized on the membrane can be released in a sustained manner for continuous stimulation of the cells attached on the membrane surface. Figure 1 illustrates the schematic diagrams of the membrane with the immobilization of single VEGF, single laminin, and dual VEGF and laminin [successive binding of VEGF and laminin]. The SMC differentiation behavior of MDSCs on the membranes immobilized with single or dual bioactive molecules was compared by DNA quantification, RT-PCR, and immunocytochemical analyses.

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  • Polymer(Korea) 폴리머
  • Frequency : Bimonthly(odd)
    ISSN 0379-153X(Print)
    ISSN 2234-8077(Online)
    Abbr. Polym. Korea
  • 2023 Impact Factor : 0.4
  • Indexed in SCIE

This Article

  • 2018; 42(5): 866-873

    Published online Sep 25, 2018

  • 10.7317/pk.2018.42.5.866
  • Received on Apr 16, 2018
  • Revised on May 2, 2018
  • Accepted on May 3, 2018

Correspondence to

  • Jin Ho Lee

  • Department of Advanced Materials and Chemical Engineering, Hannam University, Daejeon 34054, Korea

  • E-mail: jhlee@hnu.kr
  • ORCID:
    0000-0002-1528-3416