Publications by authors named "Young Min Ju"

23 Publications

  • Page 1 of 1

Synergistic effect of CNTF and GDNF on directed neurite growth in chick embryo dorsal root ganglia.

PLoS One 2020 5;15(10):e0240235. Epub 2020 Oct 5.

Wake Forest Institute for Regenerative Medicine, Winston Salem, NC, United States of America.

It is often critical to improve the limited regenerative capacity of the peripheral nerves and direct neural growth towards specific targets, such as surgically implanted bioengineered constructs. One approach to accomplish this goal is to use extrinsic neurotrophic factors. The candidate factors first need to be identified and characterized in in vitro tests for their ability to direct the neurite growth. Here, we present a simple guidance assay that allows to assess the chemotactic effect of signaling molecules on the growth of neuronal processes from dorsal root ganglia (DRG) using only standard tissue culture materials. We used this technique to quantitatively determine the combined and individual effects of the ciliary neurotrophic factor (CNTF) and glial cell line-derived neurotrophic factor (GDNF) on neurite outgrowth. We demonstrated that these two neurotrophic factors, when applied in a 1:1 combination, but not individually, induced directed growth of neuronal processes towards the source of the gradient. This chemotactic effect persists without significant changes over a wide (10-fold) concentration range. Moreover, we demonstrated that other, more general growth parameters that do not evaluate growth in a specific direction (such as, neurite length and trajectory) were differentially affected by the concentration of the CNTF/GNDF mixture. Furthermore, GDNF, when applied individually, did not have any chemotactic effect, but caused significant neurite elongation and an increase in the number of neurites per ganglion.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0240235PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7535060PMC
December 2020

Self-Assembling Peptide Solution Accelerates Hemostasis.

Adv Wound Care (New Rochelle) 2021 04 10;10(4):191-203. Epub 2020 Sep 10.

Bio-Materials and Technology Lab, Grain Science and Industry, Kansas State University, Manhattan, Kansas, USA.

One of the leading causes of death following traumatic injury is exsanguination. Biological material-based hemostatic agents such as fibrin, thrombin, and albumin have a high risk for causing infection. Synthetic peptide-based hemostatic agents offer an attractive alternative. The objective of this study is to explore the potential of h9e peptide as an effective hemostatic agent in both and models. blood coagulation kinetics in the presence of h9e peptide was determined as a function of gelation time using a dynamic rheometer. hemostatic effects were studied using the Wistar rat model. Results were compared to those of the commercial hemostatic product Celox™, a chitosan-based product. Adhesion of h9e peptide was evaluated using the platelet adhesion test. Biocompatibility of h9e peptide was studied using a mouse model. After h9e peptide solution was mixed with blood, gelation started immediately, increased rapidly with time, and reached more than 100 Pa within 3 s. Blood coagulation strength increased as h9e peptide wt% concentration increased. In the rat model, h9e peptide solution at 5% weight concentration significantly reduced both bleeding time and blood loss, outperforming Celox. Preliminary pathological studies indicate that h9e peptide solution is biocompatible and did not have negative effects when injected subcutaneously in a mouse model. For the first time, h9e peptide was found to have highly efficient hemostatic effects by forming nanoweb-like structures, which act as a preliminary thrombus and a surface to arrest bleeding 82% faster compared to the commercial hemostatic agent Celox. This study demonstrates that h9e peptide is a promising hemostatic biomaterial, not only because of its greater hemostatic effect than commercial product Celox but also because of its excellent biocompatibility based on the mouse model study.
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http://dx.doi.org/10.1089/wound.2019.1109DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7906870PMC
April 2021

Structure establishment of three-dimensional (3D) cell culture printing model for bladder cancer.

PLoS One 2019 22;14(10):e0223689. Epub 2019 Oct 22.

Department of Urology, College of Medicine, Chung-Ang University, Seoul, Republic of Korea.

Purpose: Two-dimensional (2D) cell culture is a valuable method for cell-based research but can provide unpredictable, misleading data about in vivo responses. In this study, we created a three-dimensional (3D) cell culture environment to mimic tumor characteristics and cell-cell interactions to better characterize the tumor formation response to chemotherapy.

Materials And Methods: We fabricated the 3D cell culture samples using a 3D cell bio printer and the bladder cancer cell line 5637. T24 cells were used for 2D cell culture. Then, rapamycin and Bacillus Calmette-Guérin (BCG) were used to examine their cancer inhibition effects using the two bladder cancer cell lines. Cell-cell interaction was measured by measuring e-cadherin and n-cadherin secreted via the epithelial-mesenchymal transition (EMT).

Results: We constructed a 3D cell scaffold using gelatin methacryloyl (GelMA) and compared cell survival in 3D and 2D cell cultures. 3D cell cultures showed higher cancer cell proliferation rates than 2D cell cultures, and the 3D cell culture environment showed higher cell-to-cell interactions through the secretion of E-cadherin and N-cadherin. Assessment of the effects of drugs for bladder cancer such as rapamycin and BCG showed that the effect in the 2D cell culture environment was more exaggerated than that in the 3D cell culture environment.

Conclusions: We fabricated 3D scaffolds with bladder cancer cells using a 3D bio printer, and the 3D scaffolds were similar to bladder cancer tissue. This technique can be used to create a cancer cell-like environment for a drug screening platform.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0223689PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6804961PMC
March 2020

A novel decellularized skeletal muscle-derived ECM scaffolding system for in situ muscle regeneration.

Methods 2020 01 3;171:77-85. Epub 2019 Jul 3.

Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC 27157, USA. Electronic address:

The cell-based tissue engineering strategies have gained attention in restoring normal tissue function after skeletal muscle injuries; however, these approaches require a donor tissue biopsy and extensive cell expansion process prior to implantation. In order to avoid this limitation, we developed a novel cell-free muscle-specific scaffolding system that consisted of a skeletal muscle-derived decellularized extracellular matrix (dECM) and a myogenic factor, insulin growth factor-1 (IGF-1). Rheological, morphological, and biological properties of this muscle-specific scaffold (IGF-1/dECM) as well as collagen and dECM scaffolds were examined. The cell viability in all scaffolds had over 90% at 1, 3, and 7 days in culture. The cell proliferation in the IGF-1/dECM was significantly increased when compared with other groups. More importantly, the IGF-1/dECM strongly supported the myogenic differentiation in the scaffold as confirmed by myosin heavy chain (MHC) immunofluorescence. We also investigated the feasibility in a rabbit tibialis anterior (TA) muscle defect model. The IGF-1/dECM had a significantly greater number of myofibers when compared to both collagen and dECM groups at 1 and 2 months after implantation. We demonstrated that this novel muscle-specific scaffolding system could effectively promote the muscle tissue regeneration in situ.
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http://dx.doi.org/10.1016/j.ymeth.2019.06.027DOI Listing
January 2020

In vitro evaluation of functionalized decellularized muscle scaffold for in situ skeletal muscle regeneration.

Biomed Mater 2019 06 5;14(4):045015. Epub 2019 Jun 5.

Current treatment options for repairing volumetric muscle loss injury involve the use of existing host tissue like muscular flaps or grafts. However, host muscle tissue may not be available and donor site morbidity, such as functional loss and volume deficiency, is often present. In this study, we developed a biofunctionalized muscle-derived decellularized extracellular matrix scaffolding system to utilize endogenous stem/progenitor cells for in situ muscle tissue regeneration. We optimized the decellularization process to enhance cellular infiltration and fabricated an insulin-like growth factor-binding protein 3 (IGFBP-3)-conjugated scaffold for controlled delivery of IGF-I. We then tested in vitro characterization including IGF-I release kinetics and cellular infiltration. In addition, we have analyzed the bioactivities of skeletal muscle cells (C2C12) to assess the indirect effect of released IGF-1 from the scaffold. The IGFBP-3 conjugated scaffolds demonstrated showed sustained release of IGF-1 and 1% SDS decellularized scaffold with IGF-1 showed higher cellular infiltration compared to control scaffolds (no conjugation). In indirect bioactivity assay, IGF-1 conjugated scaffold showed 2.1-fold increased cell activity compared to control (fresh media). Our results indicate that IGFBP-3/IGF-I conjugated scaffold has the potential to be used for in situ muscle tissue regeneration.
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http://dx.doi.org/10.1088/1748-605X/ab229dDOI Listing
June 2019

Electrospun vascular scaffold for cellularized small diameter blood vessels: A preclinical large animal study.

Acta Biomater 2017 09 19;59:58-67. Epub 2017 Jun 19.

Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC 27157, USA. Electronic address:

The strategy of vascular tissue engineering is to create a vascular substitute by combining autologous vascular cells with a tubular-shaped biodegradable scaffold. We have previously developed a novel electrospun bilayered vascular scaffold that provides proper biological and biomechanical properties as well as structural configuration. In this study, we investigated the clinical feasibility of a cellularized vascular scaffold in a preclinical large animal model. We fabricated the cellularized vascular construct with autologous endothelial progenitor cell (EPC)-derived endothelial cells (ECs) and smooth muscle cells (SMCs) followed by a pulsatile bioreactor preconditioning. This fully cellularized vascular construct was tested in a sheep carotid arterial interposition model. After preconditioning, confluent and mature EC and SMC layers in the scaffold were achieved. The cellularized constructs sustained the structural integrity with a high degree of graft patency without eliciting an inflammatory response over the course of the 6-month period in sheep. Moreover, the matured EC coverage on the lumen and a thick smooth muscle layer were formed at 6months after transplantation. We demonstrated that electrospun bilayered vascular scaffolds in conjunction with autologous vascular cells may be a clinically applicable alternative to traditional prosthetic vascular graft substitutes.

Statement Of Significance: This study demonstrates the utility of tissue engineering to provide platform technologies for rehabilitation of patients recovering from severe, devastating cardiovascular diseases. The long-term goal is to provide alternatives to vascular grafting using bioengineered blood vessels derived from an autologous cell source with a functionalized vascular scaffold. This novel bilayered vascular construct for engineering blood vessels is designed to offer "off-the-shelf" availability for clinical translation.
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http://dx.doi.org/10.1016/j.actbio.2017.06.027DOI Listing
September 2017

CD133 antibody conjugation to decellularized human heart valves intended for circulating cell capture.

Biomed Mater 2015 Sep 3;10(5):055001. Epub 2015 Sep 3.

Wake Forest Institute for Regenerative Medicine and, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC 27157, USA. Department of General Surgery, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC 27157, USA.

The long term efficacy of tissue based heart valve grafts may be limited by progressive degeneration characterized by immune mediated inflammation and calcification. To avoid this degeneration, decellularized heart valves with functionalized surfaces capable of rapid in vivo endothelialization have been developed. The aim of this study is to examine the capacity of CD133 antibody-conjugated valve tissue to capture circulating endothelial progenitor cells (EPCs). Decellularized human pulmonary valve tissue was conjugated with CD133 antibody at varying concentrations and exposed to CD133 expressing NTERA-2 cl.D1 (NT2) cells in a microflow chamber. The amount of CD133 antibody conjugated on the valve tissue surface and the number of NT2 cells captured in the presence of shear stress was measured. Both the amount of CD133 antibody conjugated to the valve leaflet surface and the number of adherent NT2 cells increased as the concentration of CD133 antibody present in the surface immobilization procedure increased. The data presented in this study support the hypothesis that the rate of CD133(+) cell adhesion in the presence of shear stress to decellularized heart valve tissue functionalized by CD133 antibody conjugation increases as the quantity of CD133 antibody conjugated to the tissue surface increases.
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http://dx.doi.org/10.1088/1748-6041/10/5/055001DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5104175PMC
September 2015

A novel tissue-engineered trachea with a mechanical behavior similar to native trachea.

Biomaterials 2015 Sep 23;62:106-15. Epub 2015 May 23.

Department of Mechanical Engineering, Pohang University of Science and Technology (POSTECH), 77 Cheongam-ro, Nam-gu, Pohang, Gyeongbuk, 790-784, South Korea. Electronic address:

A novel tissue-engineered trachea was developed with appropriate mechanical behavior and substantial regeneration of tracheal cartilage. We designed hollow bellows scaffold as a framework of a tissue-engineered trachea and demonstrated a reliable method for three-dimensional (3D) printing of monolithic bellows scaffold. We also functionalized gelatin sponge to allow sustained release of TGF-β1 for stimulating tracheal cartilage regeneration and confirmed that functionalized gelatin sponge induces cartilaginous tissue formation in vitro. A tissue-engineered trachea was then created by assembling chondrocytes-seeded functionalized gelatin sponges into the grooves of bellows scaffold and it showed very similar mechanical behavior to that of native trachea along with substantial regeneration of tracheal cartilage in vivo. The tissue-engineered trachea developed here represents a novel concept of tracheal substitute with appropriate mechanical behavior similar to native trachea for use in reconstruction of tracheal stenosis.
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http://dx.doi.org/10.1016/j.biomaterials.2015.05.008DOI Listing
September 2015

Engineered small diameter vascular grafts by combining cell sheet engineering and electrospinning technology.

Acta Biomater 2015 Apr 30;16:14-22. Epub 2015 Jan 30.

Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC 27157, USA.

Tissue engineering offers an attractive approach to creating functional small-diameter (<5mm) blood vessels by combining autologous cells with a natural and/or synthetic scaffold under suitable culture conditions, which results in a tubular construct that can be implanted in vivo. We have previously developed a vascular scaffold fabricated by electrospinning poly(ε-caprolactone) (PCL) and type I collagen that mimics the structural and biomechanical properties of native vessels. In this study, we investigated whether a smooth muscle cell (SMC) sheet could be combined with the electrospun vascular scaffolds to produce a more mature smooth muscle layer as compared to the conventional cell seeding method. The pre-fabricated SMC sheet, wrapped around the vascular scaffold, provided high cell seeding efficiency (approx. 100%) and a mature smooth muscle layer that expressed strong cell-to-cell junction, connexin 43 (CX43), and contractile proteins, α smooth muscle actin (α-SMA) and myosin light chain kinase (MLCK). Moreover, bioreactor-associated preconditioning of the SMC sheet-combined vascular scaffold maintained high cell viability (95.9 ± 2.7%) and phenotypes and improved cellular infiltration and mechanical properties (35.7% of tensile strength, 47.5% of elasticity, and 113.2% of elongation at break).
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http://dx.doi.org/10.1016/j.actbio.2015.01.030DOI Listing
April 2015

In situ regeneration of skeletal muscle tissue through host cell recruitment.

Acta Biomater 2014 Oct 20;10(10):4332-9. Epub 2014 Jun 20.

Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC 27157, USA. Electronic address:

Standard reconstructive procedures for restoring normal function after skeletal muscle defects involve the use of existing host tissues such as muscular flaps. In many instances, this approach is not feasible and delays the rehabilitation process and restoration of tissue function. Currently, cell-based tissue engineering strategies have been used for reconstruction; however, donor tissue biopsy and ex vivo cell manipulation are required prior to implantation. The present study aimed to overcome these limitations by demonstrating mobilization of muscle cells into a target-specific site for in situ muscle regeneration. First, we investigated whether host muscle cells could be mobilized into an implanted scaffold. Poly(l-lactic acid) (PLLA) scaffolds were implanted in the tibialis anterior (TA) muscle of rats, and the retrieved scaffolds were characterized by examining host cell infiltration in the scaffolds. The host cell infiltrates, including Pax7+ cells, gradually increased with time. Second, we demonstrated that host muscle cells could be enriched by a myogenic factor released from the scaffolds. Gelatin-based scaffolds containing a myogenic factor were implanted in the TA muscle of rats, and the Pax7+ cell infiltration and newly formed muscle fibers were examined. By the second week after implantation, the Pax7+ cell infiltrates and muscle formation were significantly accelerated within the scaffolds containing insulin-like growth factor 1 (IGF-1). Our data suggest an ability of host stem cells to be recruited into the scaffolds with the capability of differentiating to muscle cells. In addition, the myogenic factor effectively promoted host cell recruitment, which resulted in accelerating muscle regeneration in situ.
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http://dx.doi.org/10.1016/j.actbio.2014.06.022DOI Listing
October 2014

Diaphragmatic muscle reconstruction with an aligned electrospun poly(ε-caprolactone)/collagen hybrid scaffold.

Biomaterials 2013 Nov 6;34(33):8235-40. Epub 2013 Aug 6.

Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Medical Center Boulevard, Winston Salem, NC 27157, USA.

Large diaphragmatic muscle defects in congenital diaphragmatic hernia (CDH) are reconstructed by prosthetic materials or autologous grafts, which are associated with high complications and reherniation. In this study we examined the feasibility of using aligned electrospun poly(ε-caprolactone) (PCL)/collagen hybrid scaffolds for diaphragmatic muscle reconstruction. The hybrid scaffolds were implanted into a central left hemi-diaphragmatic defect (approximately 70% of the diaphragmatic tissue on the left side) in rats. Radiographic and magnetic resonance imaging (MRI) analyses showed no evidence of herniation or retraction up to 6 months after implantation. Histological and immunohistochemical evaluations revealed ingrowth of muscle tissue into the scaffolds. The mechanical properties of the retrieved diaphragmatic scaffolds were similar to those of normal diaphragm at the designated time points. Our results show that the aligned electrospun hybrid scaffolds allowed muscle cell migration and tissue formation. The aligned scaffolds may provide implantable functional muscle tissues for patients with diaphragmatic muscle defects.
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http://dx.doi.org/10.1016/j.biomaterials.2013.07.057DOI Listing
November 2013

Functional recovery of denervated muscle by neurotization using nerve guidance channels.

J Tissue Eng Regen Med 2015 Jul 12;9(7):838-46. Epub 2013 Feb 12.

Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Winston-Salem, NC, USA.

Tissue-engineered muscle has been proposed as a means of repairing volumetric muscle defects to restore anatomical and functional recovery. We have previously demonstrated that denervated muscle, which is analogous to engineered muscle construct, can be reinnervated by direct transplantation of host nerve (neurotization) in a rat model. However, the use of this approach is not possible if the length of host nerve is inadequate and cannot be mobilized to the insertion site of the engineered muscle. In this study we investigated whether neurotization coupled with nerve guidance channels would increase the regeneration of neuromuscular junctions (NMJs) in completely denervated muscle and encourage neurofunctional recovery. Seventy-two Lewis rats were evaluated in three groups, a normal control group (n = 8), a denervated group (n = 32) and a neurotization coupled with nerve guidance group (n = 32). Neurofunctional behaviour and histological evaluations were performed at 4, 8, 12 and 20 weeks postoperatively. Extensor postural thrust (EPT) and compound muscle action potential (CMAP) amplitude were significantly improved in the nerve guidance group when compared with the denervated group, even though these values were different from those of the normal control group at 20 weeks postoperation. Regeneration of axons and NMJs was demonstrated histologically in the nerve guidance group. Neurotization coupled with nerve guidance channels leads to regeneration of axons and NMJs in completely denervated muscle. To our knowledge, this is the first report to show that nerve guidance can allow re-innervation in denervated muscle containing long-gap nerve injuries.
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http://dx.doi.org/10.1002/term.1696DOI Listing
July 2015

In vitro osteogenic differentiation of human amniotic fluid-derived stem cells on a poly(lactide-co-glycolide) (PLGA)-bladder submucosa matrix (BSM) composite scaffold for bone tissue engineering.

Biomed Mater 2013 Feb 25;8(1):014107. Epub 2013 Jan 25.

Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC 27157, USA.

Stem cells have become an important component of tissue regeneration, as they are able to differentiate into various cell types if guided appropriately. It is well known that cellular differentiation is greatly influenced by the surrounding microenvironment. We have developed a composite scaffold system using a collagen matrix derived from porcine bladder submucosa matrix (BSM) and poly(lactide-co-glycolide) (PLGA). In this study, we investigated whether a composite scaffold composed of naturally derived matrix combined with synthetic polymers would provide a microenvironment to facilitate the induction of osteogenic differentiation. We first showed that human amniotic fluid-derived stem cells (hAFSCs) adhered to the composite scaffolds and proliferated over time. We also showed that the composite scaffolds facilitated the differentiation of hAFSCs into an osteogenic lineage. The expression of osteogenic genes, including RUNX2, osteopontin (OPN) and osteocalcin (OCN) was upregulated in cells cultured on the composite scaffolds incubated in the osteogenic medium compared with ones without. Increased alkaline phosphatase (ALP) activity and calcium content indicates that hAFSCs seeded on 3D porous BSM-PLGA composite scaffolds resulted in higher mineralization rates as the duration of induction increased. This was also evidenced by the mineralized matrix within the scaffolds. The composite scaffold system provides a proper microenvironment that can facilitate osteogenic differentiation of AFSCs. This scaffold system may be a good candidate material for bone tissue engineering.
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http://dx.doi.org/10.1088/1748-6041/8/1/014107DOI Listing
February 2013

End-to-side neurorrhaphy using an electrospun PCL/collagen nerve conduit for complex peripheral motor nerve regeneration.

Biomaterials 2012 Dec 19;33(35):9027-36. Epub 2012 Sep 19.

Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC 27157, USA.

In cases of complex neuromuscular defects, finding the proximal stump of a transected nerve in order to restore innervation to damaged muscle is often impossible. In this study we investigated whether a neighboring uninjured nerve could serve as a source of innervation of denervated damaged muscle through a biomaterial-based nerve conduit while preserving the uninjured nerve function. Tubular nerve conduits were fabricated by electrospinning a polymer blend consisting of poly(ε-caprolactone) (PCL) and type I collagen. Using a rat model of common peroneal injury, the proximal end of the nerve conduit was connected to the side of the adjacent uninjured tibial branch (TB) of the sciatic nerve after partial axotomy, and the distal end of the conduit was connected to the distal stump of the common peroneal nerve (CPN). The axonal continuity recovered through the nerve conduit at 8 weeks after surgery. Recovery of denervated muscle function was achieved, and simultaneously, the donor muscle, which was innervated by the axotomized TB also recovered at 20 weeks after surgery. Therefore, this end-to-side neurorrhaphy (ETS) technique using the electrospun PCL/collagen conduit appears to be clinically feasible and would be a useful alternative in instances where autologous nerve grafts or an adequate proximal nerve stump is unavailable.
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http://dx.doi.org/10.1016/j.biomaterials.2012.09.008DOI Listing
December 2012

Combined systemic and local delivery of stem cell inducing/recruiting factors for in situ tissue regeneration.

FASEB J 2012 Jan 30;26(1):158-68. Epub 2011 Sep 30.

Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, Winston-Salem, NC 27157, USA.

Whereas the conventional tissue engineering strategy involves the use of scaffolds combined with appropriate cell types to restore normal functions, the concept of in situ tissue regeneration uses host responses to a target-specific scaffold to mobilize host cells to a site of injury without the need for cell seeding. For this purpose, local delivery of bioactive molecules from scaffolds has been generally used. However, this approach has limited stem cell recruitment into the implants. Thus, we developed a combination of systemic delivery of substance P (SP) and local release of stromal-derived factor-1α (SDF-1α) from an implant. In this study, we examined whether this combined system would significantly enhance recruitment of host stem cells into the implants. Flow cytometry and immunohistochemistry for CD29/CD45, CD146/α-smooth muscle actin, and c-kit demonstrated that this system significantly increased the number of stem cell-like cells within the implants when compared with other systems. In vitro culture of the cells that had infiltrated into the scaffolds from the combined system confirmed that host stem cells were recruited into these implants and indicated that they were capable of differentiation into multiple lineages. These results indicate that this combined system may lead to more efficient tissue regeneration.
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http://dx.doi.org/10.1096/fj.11-182998DOI Listing
January 2012

Bilayered scaffold for engineering cellularized blood vessels.

Biomaterials 2010 May 25;31(15):4313-21. Epub 2010 Feb 25.

Wake Forest Institute for Regenerative Medicine, Wake Forest University Health Sciences, Medical Center Boulevard, Winston-Salem, NC 27157, USA.

Vascular scaffolds fabricated by electrospinning poly(epsilon-caprolactone) (PCL) and collagen have been designed to provide adequate structural support as well as a favorable adhesion substrate for vascular cells. However, the presence of small-sized pores limits the efficacy of smooth muscle cells (SMC) seeding, as these cells could not adequately infiltrate into the scaffolds. To overcome this challenge, we developed a bilayered scaffolding system that provides different pore sizes to facilitate adequate cellular interactions. Based on the fact that pore size increases with the increase in fiber diameter, four different fiber diameters (ranging 0.27-4.45 mum) were fabricated by electrospinning with controlled parameters. The fabricated scaffolds were examined by evaluating cellular interactions, and the mechanical properties were measured. Endothelial cells (EC) seeded on nanoscaled fibers showed enhanced cellular orientation and focal adhesion. Conversely, fabrication of a larger fiber diameter improved SMC infiltration into the scaffolds. To incorporate both of these properties into a scaffold, bilayered vascular scaffolds were produced. The inner layer yielded small diameter fibers and the outer layer provided large diameter fibers. We show that the bilayered scaffolds permit EC adhesion on the lumen and SMC infiltration into the outer layer. This study suggests that the use of bilayered scaffolds may lead to improved vessel formation.
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http://dx.doi.org/10.1016/j.biomaterials.2010.02.002DOI Listing
May 2010

A dexamethasone-loaded PLGA microspheres/collagen scaffold composite for implantable glucose sensors.

J Biomed Mater Res A 2010 Apr;93(1):200-10

Department of Chemical & Biomedical Engineering, University of South Florida, Tampa, FL 33620-5350, USA.

We have developed a new dexamethasone (Dex)-loaded poly(lactic-co-glycolic acid) microspheres/porous collagen scaffold composite for implantable glucose sensors. The scaffolds were fabricated around the sensing element of the sensors and crosslinked using nordihydroguaiaretic acid (NDGA). The microspheres containing Dex were incorporated into the NDGA-crosslinked collagen scaffold by dipping in microsphere suspension in either water or Pluronic. The loading efficiencies of Dex in the microspheres and the scaffold were determined using high performance liquid chromatography. The microspheres/scaffold composite fabricated using microspheres in the hydrogel solution had a better loading efficiency than when microspheres were in water suspension. The composite fabricated using the hydrogel also showed a slower and more sustained drug release than the standard microspheres in vitro during a 4 week study and did not significantly affect the function of the sensors in vitro. The sensors with the composite that were still functional retained above 50% of their original sensitivity at 2 weeks. Histology showed that the inflammatory response to the Dex-loaded composite was much lower than for the control scaffold at 2 and 4 weeks after implantation. The Dex-loaded composite system might be useful to reduce inflammation to implanted glucose sensors and therefore extend their function and lifetime.
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http://dx.doi.org/10.1002/jbm.a.32512DOI Listing
April 2010

A novel porous collagen scaffold around an implantable biosensor for improving biocompatibility. II. Long-term in vitro/in vivo sensitivity characteristics of sensors with NDGA- or GA-crosslinked collagen scaffolds.

J Biomed Mater Res A 2010 Feb;92(2):650-8

Biomedical Engineering Program, University of South Florida, 4202 E. Fowler Avenue, ENB 118, Tampa, Florida 33620-5350, USA.

We have developed a new 3D porous and biostable collagen scaffold for implantable glucose sensors. The scaffolds were fabricated around the sensors and crosslinked using nordihydroguaiaretic acid (NDGA) or glutaraldehyde (GA) to enhance physical and biological stability. The effect of the scaffolds on sensor function and biocompatibility was examined during long-term (>or=28 days) in vitro and in vivo experiments and compared with control bare sensors. To evaluate the effect of the sensor length on micromotion and sensor function, we also fabricated short and long sensors. 3D porous scaffold application around glucose sensors did not significantly affect the long-term in vitro sensitivity of the sensors. The scaffolds, crosslinked by either NDGA or GA, remained stable around the sensors during the 4 week in vitro study. In the long-term in vivo study, the sensitivity of the short sensors was higher than the sensitivity of long sensors presumably because of less micromotion in the subcutis of the rats. The sensors with NDGA-crosslinked scaffolds had a higher sensitivity than the sensors with GA-crosslinked scaffolds. Histological examination showed that NDGA-crosslinked scaffolds retained their physical structure with reduced inflammation when compared with the GA-crosslinked scaffolds. Therefore, the application of NDGA-crosslinked collagen scaffolds might be a good method for enhancing the function and lifetime of implantable biosensors by minimizing the in vivo foreign body response.
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http://dx.doi.org/10.1002/jbm.a.32400DOI Listing
February 2010

Use of hydrogel coating to improve the performance of implanted glucose sensors.

Biosens Bioelectron 2008 Mar 29;23(8):1278-84. Epub 2007 Nov 29.

Department of Chemical Engineering, University of South Florida, 4202 E. Fowler Avenue, ENB 118, Tampa, FL 33620-5350, UK.

In order to protect implanted glucose sensors from biofouling, novel hydrogels (146-217% water by mass) were developed based on a copolymer of hydroxyethyl methacrylate (HEMA) and 2,3-dihydroxypropyl methacrylate (DHPMA). The porosity and mechanical properties of the hydrogels were improved using N-vinyl-2-pyrrolidinone (VP) and ethyleneglycol dimethacrylate (EGDMA). The results of SEM and DSC FT-IT analyses showed that the hydrogel (VP30) produced from a monomeric mixture of 34.5% HEMA, 34.5% DHPMA, 30% VP and 1% EDGMA (mol%) had an excellent pore structure, high water content at swelling equilibrium (W eq=166% by mass) and acceptable mechanical properties. Two kinds of VP30-coated sensors, Pt/GOx/VP30 and Pt/GOx/epoxy-polyurethane (EPU)/VP30 sensors were examined in glucose solutions during a period of 4 weeks. The Pt/GOx/VP30 sensors produced large response currents but the response linearity was poor. Therefore, further studies were focused on the Pt/GOx/EPU/VP30 sensors. With a diffusion-limiting epoxy-polyurethane membrane, the linearity was improved (2-30 mM) and the response time was within 5 min. Eight Pt/GOx/EPU/VP30 sensors were subcutaneously implanted in rats and tested once per week over 4 weeks. All of the implanted sensors kept functioning for at least 21 days and 3 out of 8 sensors still functioned at day 28. Histology revealed that the fibrous capsules surrounding hydrogel-coated sensors were thinner than those surrounding Pt/GOx/EPU sensors after 28 days of implantation.
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http://dx.doi.org/10.1016/j.bios.2007.11.010DOI Listing
March 2008

A novel porous collagen scaffold around an implantable biosensor for improving biocompatibility. I. In vitro/in vivo stability of the scaffold and in vitro sensitivity of the glucose sensor with scaffold.

J Biomed Mater Res A 2008 Oct;87(1):136-46

Biomedical Engineering Program, University of South Florida, 4202 E. Fowler Avenue, ENB 118, Tampa, Florida 33620-5350, USA.

A new 3D porous and biostable collagen scaffold has been developed to improve the biocompatibility of implantable glucose sensors by minimizing tissue reactions while stimulating angiogenesis around the sensors. The novel collagen scaffold was crosslinked using nordihydroguaiaretic acid (NDGA) to enhance biostability. NDGA-treated collagen scaffolds were stable without physical deformation in the subcutaneous tissue of rats for 4 weeks. In contrast, glutaraldehyde (GA)-treated collagen scaffolds were extremely damaged following implantation. Both types of scaffolds (NDGA- and GA-crosslinked) were stable in vitro in the presence of collagenase with 70% retention of original weight after 4 weeks of incubation. The response current (i.e. sensitivity) of sensors with porous scaffolds was not significantly changed when compared with control sensors (no scaffold), while the response time (T(95%)) was slightly delayed after a glucose concentration increase from 5 to 15 mM. Above this range, the sensors coated with scaffolds had only a slightly lower sensitivity than the control sensors. These results indicate that we have developed a stable NDGA-crosslinked collagen scaffold for biosensors, and that the scaffold does not impair the function of our sensor. We plan to use this scaffold to enhance the function and lifetime of the implantable biosensors by providing a controlled local environment around the sensors with the help of various drugs and growth factors (dexamethasone, VEGF, PDGF).
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http://dx.doi.org/10.1002/jbm.a.31756DOI Listing
October 2008

Beneficial effect of hydrophilized porous polymer scaffolds in tissue-engineered cartilage formation.

J Biomed Mater Res B Appl Biomater 2008 Apr;85(1):252-60

Biomaterials Research Center, Korea Institute of Science and Technology, P.O. Box 131, Cheongryang, Seoul 130-650, Korea.

Three dimensional (3D) porous poly(L-lactic acid) (PLLA) scaffolds were fabricated using a modified gas foaming method whose effervescent porogens were a mixture of sodium bicarbonate and citric acid. To improve chondrocyte adhesion, the scaffolds were then hydrophilized through oxygen plasma treatment and in situ graft polymerization of acrylic acid (AA). When the physical properties of AA-grafted scaffolds were examined, the porosity and pore size were 87 approximately 93% and 100 approximately 300 microm, respectively. The pore sizes were highly dependent on the varying ratios (w/w) between porogen and polymer solution. Influenced by their pore sizes, the compressive moduli of scaffolds significantly decreased with increasing pore size. The altered surface characteristics were clearly reflected in the reduced water contact angles that meant a significant hydrophilization with the modified polymer surface. Electron spectroscopy for chemical analysis (ESCA) and time-of-flight secondary ion mass spectrometer (ToF-SIMS) also confirmed the altered surface chemistry. When chondrocytes were seeded onto the AA-grafted PLLA scaffolds, cell adhesion and proliferation were substantially improved as compared to the unmodified scaffolds. The benefit of the modified scaffolds was clear in the gene expressions of collagen type II that was significantly upregulated after 4-week culture. Safranin-O staining also identified greater glycosaminoglycan (GAG) deposition in the modified scaffold. The AA-grafted porous polymer scaffolds were effective for cell adhesion and differentiation, making them a suitable platform for tissue-engineered cartilage.
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http://dx.doi.org/10.1002/jbm.b.30943DOI Listing
April 2008

Surface modification of biodegradable electrospun nanofiber scaffolds and their interaction with fibroblasts.

J Biomater Sci Polym Ed 2007 ;18(4):369-82

Biomaterials Research Center, Korea Institute of Science and Technology, P.O. Box 131, Cheongryang, Seoul 130-650, South Korea.

Biodegradable polymers, such as poly(glycolic acid) (PGA), poly(L-lactic acid) (PLLA) and poly(lactic-co-glycolic acid) (PLGA), were dissolved individually in the proper solvents and then subjected to electrospinning process to make nanofibrous scaffolds. Their surfaces were then chemically modified using oxygen plasma treatment and in situ grafting of hydrophilic acrylic acid (AA). The fiber thickness, pore size and porosity were estimated to 200-800 nm, 2-30 microm and 94-96%, respectively, and these properties were insignificant in the PGA, PLLA and PLGA nanofibrous scaffolds. The ultimate tensile strength of PGA was about 2.5 MPa on average and that of PLGA and PLLA was less than 2 MPa. The elongation-at-break was 100-130% for the three nanofibrous scaffolds. When the surface properties of AA-grafted scaffolds were examined, higher ratios of oxygen to carbon, lower contact angles and the presence of carboxylic (-COOH) groups were identified. The properties were significantly different from those of the unmodified nanofibrous scaffolds. Fibroblasts once seeded on the scaffolds were spreading over large surface area on the AA-grafted surface as compared to the unmodified PGA, PLLA and PLGA nanofibrous scaffolds. Cultured for up to 6 days, the fibroblast proliferation was found to be much better on the surface-modified nanofibrous scaffolds. The present study showed that, with the use of plasma treatment and AA grafting, the hydrophilic functional groups could be successfully adapted on the surface of electrospun nanofibrous scaffolds. Those surface-modified scaffolds made significant improvement on cell attachment and proliferation in vitro.
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http://dx.doi.org/10.1163/156856207780424997DOI Listing
July 2007

Comparative cytotoxicity of low-osmolar nonionic and high-osmolar ionic contrast media to dog gallbladder epithelial cells.

Gastrointest Endosc 2002 Mar;55(3):382-6

Division of Gastroenterology, Asan Medical Center, 388-1 Poongnap-dong, Songpa-gu, Seoul 138-736, South Korea.

Background: Most studies of the adverse effects of x-ray contrast media used in ERCP have focused on post-ERCP pancreatitis. However, the biliary epithelial cells are also exposed to contrast media during ERCP and acute cholangitis is also a serious complication of ERCP. The present study compared the cytotoxicity with gallbladder epithelial cells of ionic and nonionic contrast agents.

Methods: A high-osmolar ionic contrast agent (meglumine ioxithalamate) and a low-osmolar nonionic contrast agent (iopromide) were tested. Monolayer cell cultures of dog gallbladder epithelial cells were used. The cells were exposed to the 2 contrast agents with increasing iodine concentration and osmolality for 2 days. Cell number, S-phase fraction, aneuploidy, and supernatant LDH activities were measured each day.

Results: Cell growth was more severely inhibited by ioxithalamate than iopromide (p < 0.05) and strongly dependent on the osmolality of contrast agent. The cytostatic effect estimated by S-phase fraction was more pronounced for ioxithalamate. Chromosomal damage determined by aneuploidy was more frequently detected with ioxithalamate.

Conclusions: High-osmolar ionic contrast media are more cytotoxic than low-osmolar nonionic contrast media to gallbladder epithelial cells. Animal and clinical studies are needed to estimate the clinical implications of these findings.
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http://dx.doi.org/10.1067/mge.2002.121595DOI Listing
March 2002