Direct and Enhanced Coating method of Extracellular Matrix on scaffold using magnesium hydroxide and decellulization for Renal Tissue Regeneration

김윤아,전소영,유승권,권태균,한동근,정윤기 한국공업화학회 2018 한국공업화학회 연구논문 초록집 Vol.2018 No.0

Renal diseases have been recognized as major health issues. In this study, fibroblast-derived extracellular matrix (FDM) was directly coated on three-dimensional porous poly(lactide-co-glycolide) (PLGA) scaffold incorporated with magnesium hydroxide (MH). Optimized decellularization of confluent cells and MH effect resulted in the formation of a well-adhered and high-quality matrix. PLGA scaffold with FDM layer demonstrated significantly increased adhesion and viability of urine derived nephron progenitor cells. For PLGA/MH/FDM scaffolds, enhanced reconstruction of glomerulus was confirmed in a partially nephrectomized mice model with. These results suggest that surface coating of FDM layer using MH would be effective for renal tissue regeneration

Biomimetic Porous PLGA Scaffolds Incorporating Decellularized Extracellular Matrix for Kidney Tissue Regeneration

Lih, Eugene,Park, Ki Wan,Chun, So Young,Kim, Hyuncheol,Kwon, Tae Gyun,Joung, Yoon Ki,Han, Dong Keun American Chemical Society 2016 ACS APPLIED MATERIALS & INTERFACES Vol.8 No.33

<P>Chronic kidney disease is now recognized as a major health problem, but current therapies including dialysis and renal replacement have many limitations. Consequently, biodegradable scaffolds to help repairing injured tissue are emerging as a promising approach in the field of kidney tissue engineering. Poly(lactic-co-glycolic acid) (PLGA) is a useful biomedical material, but its insufficient biocompatibility caused a reduction in cell behavior and function. In this work, we developed the kidney-derived extracellular matrix (ECM) incorporated PLGA scaffolds as a cell supporting material for kidney tissue regeneration. Biomimetic PLGA scaffolds (PLGA/ECM) with different ECM concentrations were prepared by an ice particle leaching method, and their physicochemical and mechanical properties were characterized through various analyses. The proliferation of renal cortical epithelial cells on the PLGA/ECM scaffolds increased with an increase in ECM concentrations (0.2, 1, 5, and 10%) in scaffolds. The PLGA scaffold containing 10% of ECM has been shown to be an effective matrix for the repair and reconstitution of glomerulus and blood vessels in partially nephrectomized mice in vivo, compared with only PLGA control. These results suggest that not only can the tissue-engineering techniques be an effective alternative method for treatment of kidney diseases, but also the ECM incorporated PLGA scaffolds could be promising materials for biomedical applications including tissue engineered scaffolds and biodegradable implants.</P>

Mechanically Reinforced Extracellular Matrix Scaffold for Application of Cartilage Tissue Engineering

오현주,김순희,조재호,박상혁,민병현 한국조직공학과 재생의학회 2018 조직공학과 재생의학 Vol.15 No.3

Scaffolds with cartilage-like environment and suitable physical properties are critical for tissue-engineered cartilage repair. In this study, decellularized porcine cartilage-derived extracellular matrix (ECM) was utilized to fabricate ECM scaffolds. Mechanically reinforced ECM scaffolds were developed by combining salt-leaching and crosslinking for cartilage repair. The developed scaffolds were investigated with respect to their physicochemical properties and their cartilage tissue formation ability. The mechanically reinforced ECM scaffold showed similar mechanical strength to that of synthetic PLGA scaffold and expressed higher levels of cartilage-specific markers compared to those expressed by the ECM scaffold prepared by simple freeze-drying. These results demonstrated that the physical properties of ECM-derived scaffolds could be influenced by fabrication method, which provides suitable environments for the growth of chondrocytes. By extension, this study suggests a promising approach of natural biomaterials in cartilage tissue engineering.

Refunctionalization of Decellularized Organ Scaffold of Pancreas by Recellularization: Whole Organ Regeneration into Functional Pancreas

Uday Chandrika K.,Tripathi Rekha,Kameshwari Y.,Rangaraj Nandini,Mahesh Kumar J.,Singh Shashi 한국조직공학과 재생의학회 2021 조직공학과 재생의학 Vol.18 No.1

BACKGROUND: Tissue engineering centers on creating a niche similar to the natural one, with a purpose of developing an organ construct. A natural scaffold can replace none while creating a scaffold unique to each tissue in composition, architecture and cues that regulate the character of cells. METHODS: Whole pancreas from mouse was decellularized using detergent and enzymes, followed by recellularizing with MSC from human placenta. This construct was transplanted in streptozotocin induced diabetic mice. Histopathology of both decellularized and recellularized transplanted pancreas and qPCR analysis were performed to assess its recovery. RESULTS: Decellularization removes the cells leaving behind extracellular matrix rich natural scaffold. After reseeding with mesenchymal stem cells, these cells differentiate into pancreas specific cells. Upon transplantation in streptozotocin induced diabetic mice, this organ was capable of restoring its histomorphology and functioning. Restoration of endocrine (islets), the exocrine region (acinar) and vascular network was seen in transplanted pancreas. The process of functional recovery of endocrine system took about 20 days when the mice start showing blood glucose reduction, though none achieved gluconormalization. CONCLUSION: Natural decellularized scaffolds of soft organs can be refunctionalized using recipient’s mesenchymal stem cells to restore structure and function; and counter immune problems arising during transplantation.

Polymer mesh scaffold combined with cell-derived ECM for osteogenesis of human mesenchymal stem cells

노용권,Ping Du,김인걸,Jaehoon Ko,Seong Who Kim,Kwideok Park 한국생체재료학회 2016 생체재료학회지 Vol.20 No.2

Background: Tissue-engineered scaffold should mimic the structure and biological function of the extracellular matrix and have mechanically supportive properties for tissue regeneration. In this study, we utilized a PLGA/PLA mesh scaffold, coated with cell-derived extracellular matrix (CDM) and assessed its potential as an osteogenic microenvironment for human umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs). CDM was obtained by decellularization of in vitro-cultured type I collagen overexpressing (Col I -293 T-DK) cells. Test groups are mesh itself (control), fibronectin-coated (FN-mesh), and CDM-coated mesh scaffold (CDM-mesh). CDM was then solubilized and used for scaffold coating. Results: CDM was successfully collected and applied to mesh scaffolds. The presence of CDM was confirmed via SEM and FN immunofluorescence. After then, UCB-MSCs were seeded into the scaffolds and subjected to the induction of osteogenic differentiation for 21 days in vitro. We found that the seeded cells were viable and have better proliferation activity on CDM-mesh scaffold. In addition, when osteogenic differentiation of UCB-MSCs was examined for up to 21 days, alkaline phosphatase (ALP) activity and osteogenic marker (COL I, ALP, osteocalcin, bone sialoprotein) expression were significantly improved with UCB-MSCs when cultured in the CDM-mesh scaffold compared to the control and FN-mesh. Conclusion: Polymer mesh scaffold incorporated with CDM can provide UCB-MSCs with a better microenvironment for osteogenesis in vitro.

Cellular surface-modified scaffolds for delivery of nephron progenitor cell for renal tissue regeneration

( Md Lemon Hasan ),김윤아,( So Young Chun ),( Seung Kwon You ),( Tae Gyun Kwon ),( Dong Keun Han ),( Won-gun Koh ),정윤기 한국공업화학회 2019 한국공업화학회 연구논문 초록집 Vol.2019 No.1

Reneal tissue regeneration is expected to be a promising strategy, in context of currently limmited treatment and organ donation for cronic kideny disese. In recent year, extracellular matrix (ECM) derived scaffold, owing to its highly biocompatiblity and ability to guid renel tissue regeneration, have been widely investigated to treat kidney regeneration. In our previous study, magnesium hydroxide nanoparticles incorporated three dimensional porous PLGA scaffold was directly coated with fibroblast derived extracellular matrix. Inspired from previous study, here we have prepared nephron progenitor cell containing ECM surface- modified scaffolds for renal tissue regeneration. In in vitro cell studies with NPCs, modified scaffold showed improved cell viability and significantly increased growth factor release in respective time.

Cartilage engineering using cell-derived extracellular matrix scaffold in vitro

Jin, Cheng Zhe,Choi, Byung Hyune,Park, So Ra,Min, Byoung-Hyun Wiley Subscription Services, Inc., A Wiley Company 2010 Journal of biomedical materials research. Part A Vol.a92 No.4

<P>A cell-derived extracellular matrix (ECM) scaffold was constructed using cultured porcine chondrocytes via a freeze-drying method, and its ability to promote cartilage formation was evaluated in vitro. Scanning electron microscope (SEM) revealed that the scaffold had highly uniform porous microstructure. Then, rabbit chondrocytes were seeded dynamically on ECM scaffold and cultured for 2 days, 1, 2, and 4 weeks in vitro for analysis. Polyglycolic acid (PGA) scaffold was used as a control. On gross observation of neocartilage tissue, a silvery white cartilage-like tissue was observed after 1 week of culture in ECM scaffold, while similar morphology was seen only after 4 weeks in PGA scaffold. The volume of neocartilage-like tissue was significantly increased in both ECM and PGA groups. The compressive strength was gradually increased with time in ECM group, while gradually decreased in PGA group. DNA, glycosaminoglycan (GAG) and collagen contents also increased gradually with time in both groups, but showed more significant increase in ECM group. Histological staining for GAG (Safranin O staining) and type II collagen (immunohistochemistry) showed sustained accumulation of ECM molecules with time, which gradually and uniformly filled porous space in ECM scaffold. On the contrary, they accumulated only at the peripheral area of PGA scaffold. These results suggest that a novel cell-derived ECM scaffold can provide a promising environment for generating a high quality cartilage in vitro. © 2009 Wiley Periodicals, Inc. J Biomed Mater Res, 2010</P>

Bio-inspired configurable multiscale extracellular matrix-like structures for functional alignment and guided orientation of cells

김다운,김장호 한국공업화학회 2018 한국공업화학회 연구논문 초록집 Vol.2018 No.0

Inspired by the hierarchically organized protein fibers in extracellular matrix (ECM) as well as the physiological importance of multiscale topography, we developed a simple method for the design and manipulation of precisely controllable multiscale hierarchical structures. In this study, we approached a conceptual platform that can mimic the hierarchically multi-scale topographical and orientation cues of the ECM for controlling cell structure and function. Using our platforms, the structures and orientations of fibroblast cells were greatly influenced by the nanotopography, rather than the micro-topography. We also proposed a new approach that enables the generation of native ECM having nanofibers in specific three-dimensional configurations by culturing fibroblast cells on the multi-scale substrata. We suggest that our methodology could be used as efficient strategies for the design and manipulation of various functional platforms for advanced regenerative medicine applications.

PLGA/피브린 지지체가 늑연골 세포의 부착과 성장에 미치는 영향

장지은(Ji Eun Jang),이동원(Dong Won Lee),강길선(Gil Son Khang),송정은(Jeong Eun Song),이유정(Yu Jung Lee),이윤미(Yun Me Lee),조선아(Sun Ah Cho) 한국고분자학회 2013 폴리머 Vol.37 No.2

Poly(lactide-co-glycolic acid)(PLGA)는 좋은 기계적 성질과 생분해성으로 약물전달시스템 또는 조직공학적으로 널리 이용되고 있으나 낮은 세포 부착률을 가지고 있어 피브린을 첨가하여 이를 보완하고자 하였다. 본 연구에서 사용된 지지체는 트롬빈과 피브리노겐, 그리고 세포을 혼합시킨 후 PLGA 지지체 위에 도포시켜 제조하였다. 세포의 부착 및 증식률을 측정하고자 PLGA/피브린 지지체에 늑연골 세포를 파종 후 1, 3일 및 7일 후 SEM과 MTT 분석을 통하여 측정하였으며, 세포외기질 형성에 미치는 피브린의 영향을 확인하고자 세포를 파종 후 누드마우스에 이식하여 GAG 및 콜라겐 합성의 효과를 확인하였다. 따라서 본 연구에서는 피브린이 혼합된 PLGA 지지체가 생체내·외 환경에서 세포의 부착 및 증식에 미치는 영향을 확인하고자 연구를 진행하였다. 그 결과, PLGA/피브린 지지체가 기존의 PLGA 지지체와 비교하여 탁월한 세포 성장률을 나타내는 것으로 확인하였다. Poly(lactide-co-glycolic acid) (PLGA) has been widely used in the drug delivery and tissue engineering applications because of its good mechanical strength and biodegradation profile. However, cell attachment to the scaffold is low compared with that on fibrin although cells can be attached to the polymer surface. In this study, PLGA scaffolds were soaked in cells-fibrin suspension and polymerized with dropping fibrinogen-thrombin solution. Cellular proliferation activity was observed in PLGA/fibrin-seeded costal cartilage cells (CC) on 1, 3, and 7 days using the MTT assay and SEM. The effects of fibrin on the extracellular matrix (ECM) formation were evaluated using CC cell-seeded PLGA/fibrin scaffolds. The PLGA/fibrin scaffolds elicited more production of glycosaminoglycan (GAG) and collagen than the PLGA scaffold. In this study, fibrin incorporated PLGA scaffolds were prepared to evaluate the effects of fibrin on the cell attachment and proliferation in vitro and in vivo. In this result, we confirmed that proliferation of cells in PLGA/fibrin scaffolds were better than in PLGA scaffolds. The PLGA/fibrin scaffolds provide suitable environment for growth and proliferation of costal cartilage cells.

Micro environment modified scaffolds for impanted of nephron progenitor cell for renal tissue regeneration

김윤아,전소영,유승권,권태균,한동근,고원건,정윤기 한국공업화학회 2019 한국공업화학회 연구논문 초록집 Vol.2019 No.0

Renal diseases have been recognized as major health issues because of absence of efficient treatment and lack of organ donation. In our previous study, fibroblast-derived extracellular matrix was directly coated on three-dimensional porous PLGA scaffolds incorporated with magnesium hydroxide nanoparticles for renal regeneration. Herein, cell-derived ECM surface- modified scaffolds was more contained nephron progenitor cell (NPC) than only PLGA scaffolds. The viability and growth of NPC on the scaffolds also were improved. In vivo animal study demonstrated that the further developed scaffolds facilitate the paracrine effects of NPC and restore of renal functions.