• Journal of Periodontal and Implant Science
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• v.43 no.6
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• pp.251-261
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• 2013

A paradigm shift is taking place in medicine and dentistry from using synthetic implants and tissue grafts to a tissue engineering approach that uses degradable porous three-dimensional (3D) material hydrogels integrated with cells and bioactive factors to regenerate tissues such as dental bone and other oral tissues. Hydrogels have been established as a biomaterial of choice for many years, as they offer diverse properties that make them ideal in regenerative medicine, including dental applications. Being highly biocompatible and similar to native extracellular matrix, hydrogels have emerged as ideal candidates in the design of 3D scaffolds for tissue regeneration and drug delivery applications. However, precise control over hydrogel properties, such as porosity, pore size, and pore interconnectivity, remains a challenge. Traditional techniques for creating conventional crosslinked polymers have demonstrated limited success in the formation of hydrogels with large pore size, thus limiting cellular infiltration, tissue ingrowth, vascularization, and matrix mineralization (in the case of bone) of tissue-engineered constructs. Emerging technologies have demonstrated the ability to control microarchitectural features in hydrogels such as the creation of large pore size, porosity, and pore interconnectivity, thus allowing the creation of engineered hydrogel scaffolds with a structure and function closely mimicking native tissues. In this review, we explore the various technologies available for the preparation of macroporous scaffolds and their potential applications.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.29 no.6
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• pp.374-378
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• 2003

Aim : Several injectable materials have been used in the application of osteogenic bone substitute; however, nothing has won universal acceptance. This study was performed to investigate whether chitosan-alginate gel/MSCs/BMP-2 composites are potentially injectable materials for new bone formation. Material and Methods : The composites were injected into the subcutaneous space on the dorsum of the nude mouse to investigate whether new bone would be tissue engineered in the mouse. The composites were examined histologically over a 12-week period. Results : The composites implanted in the mouse were able to tissue engineer new bone, and the newly formed bone consisted of trabecular bone and calcified bone matrix. Conclusions : The present study shows that chitosan-alginate gel/MSCs/BMP-2 composites have the potential to become real injectable materials for new bone formation.

• Polymer(Korea)
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• v.27 no.3
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• pp.226-234
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• 2003

Ipriflavone (IP), a non-hormonal isoflavone derivative, has been shown to interfere with bone remodeling by inhibiting bone resorption and stimulating bone formation. IP consistently increased the amount of Ca incorporated into the cell layer by mesenchymal stem cells (MSCs). In this study, we developed the novel IP loaded poly(L-lactide-co-glycolide) (PLGA) scaffolds for the possibility of the application of the tissue engineered bone. IP/PLGA scaffo1ds were prepared by solvent casting/salt leaching method and were characterized by porosimeter, scanning electron microscopy, determination of residual salt amount, differential scanning calorimetry, and X-ray diffractometer, respectively. IP/PLGA scaffolds were implanted into the back of athymic nude mouse to observe the effect of IP on the osteoinduction compared with control PLGA scaffo1ds. Thin sections were cut from paraffin embedded tissues and histological sections were stained H&E, von Kossa, and immunohistochemical staining for Type I collagen and osteocalcin. It can be observed that the porosity was above 91.7% and the pore size was above 101 $\mu\textrm{m}$. Control scaffo1d and IP/PLGA scaffo1ds of 50% IP were implanted on the back of athymic nude mouse to observe the effect of IP on the induction of cells proliferation for 9 weeks. The evidence of calcification, osteoblast, and osteoid from the undifferentiated stem cells in the subcutaneous sites and other soft connective tissue sites having a preponderance of stem cells has been observed. From these results, it seems that IP plays an important role for bone induction in IP/PLCA scaffolds.

• Journal of Chest Surgery
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• v.37 no.3
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• pp.220-227
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• 2004

The number of patients with coronary artery disease and peripheral vascular disease are increasing, and the need of small diameter vessel is also increasing. We developed small diameter artificial vessel and experimented in vivo. We got allogenic valve from mongrel dogs, and removed all cells from the allogenic valve. Then, we seeded autologous bone marrow cells onto the decellularized scaffold. After implantation of artificial vessel into the canine carotid artery, we performed angiography regularly. In case of vessel occlusion or at 8 weeks after operation, we euthanized dogs, and retrieved the implanted artificial vessels. Control vessels were all occluded except one (which developed aneurysmal dilatation). But autologous cell seeded vascular graft were patent by 4 weeks in one, by 6 in one and by 8 weeks in two. Histologic examination of patent vessel revealed similar structure to native artery. Tissue-engineered vascular graft manufactured with decellularized allogenic matrix and autologous bone marrow cells showed that tissue engineered graft had similar structure to native artery.

• Polymer(Korea)
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• v.28 no.5
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• pp.382-390
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• 2004

One of the significant natural bioactive materials is demineralized bone particle (DBP) whose has a powerful induce. of new bone growth. In this study, we developed the DBP loaded poly-lactide (PLA) and poly(L-lactide-co-glycolide) (PLGA) scaffolds for the possibility of the application of the tissue engineered bone. PLA/DBP and PLGA/DBP scaffolds were prepared by solvent casting/salt leaching method and were characterized by porosimeter, scanning electron microscopy. BMSCs were stimulated by osteogenic medium and characterized by histological stained Wright-Giemsa, Alizarin red, von Kossa, and alkaline phosphate activity (ALP). DBP impregnated scaffolds with BMSCs were implanted into the back of athymic nude mouse to observe the effect of DBP on the osteoinduction compared with control scaffolds. It can be observed that the porosity was above $90.2\%$ and the pore size was above 69.1$\mu$m. BMSCs could be differentiated into osteoprogenitor cells as result of wright-giemsa, alizarin red, von Kossa and ALP staining. In in vivo study, we could observed calcification region in PLA/DBP and PLGA/DBP groups, but calcification did not occur almost in control scaffolds. From these results, it seems that DBP as well as BMSCs play an important role for bone induction in PLA/DBP and PLGA/DBP scaffolds.

• Archives of Pharmacal Research
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• v.26 no.1
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• pp.76-82
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• 2003

Drug releasing porous poly($\varepsilon$-caprolactone) (PCL)-chitosan matrices were fabricated for bone regenerative therapy. Porous matrices made of biodegradable polymers have been playing a crucial role as bone substitutes and as tissue-engineered scaffolds in bone regenerative therapy. The matrices provided mechanical support for the developing tissue and enhanced tissue formation by releasing active agent in controlled manner. Chitosan was employed to enhance hydrophilicity and biocompatibility of the PCL matrices. PDGF-BB was incorporated into PCL-chitosan matrices to induce enhanced bone regeneration efficacy. PCL-chitosan matrices retained a porous structure with a 100-200 $\mu$m pore diameter that was suitable for cellular migration and osteoid ingrowth. $NaHCO_3$ as a porogen was incorporated 5% ratio to polymer weight to form highly porous scaffolds. PDGF-BB was released from PCL-chitosan matrices maintaining therapeutic concentration for 4 week. High osteoblasts attachment level and proliferation was observed from PCL-chitosan matrices. Scanning electron microscopic examination indicated that cultured osteoblasts showed round form and spread pseudopods after 1 day and showed broad cytoplasmic extension after 14 days. PCL-chitosan matrices promoted bone regeneration and PDGF-BB loaded matrices obtained enhanced bone formation in rat calvarial defect. These results suggested that the PDGF-BB releasing PCL-chitosan porous matrices may be potentially used as tissue engineering scaffolds or bone substitutes with high bone regenerative efficacy.

• Polymer(Korea)
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• v.31 no.1
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• pp.14-19
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• 2007

Demineralized bone particle (DBP) has been used as one of the powerful inducers of bone and cartilage tissue specialization. In this study, we fabricated DBP/PLGA scaffold for tissue engineered disc regeneration. We manufactured dual-structured scaffold to compose inner cylinder and outer doughnut similar to nature disc tissue. The DBP/PLGA scaffold was characterized by porosity, wettability, and water uptake ability. We isolated and cultured nucleus pulposus (NP) and annulus fibrosus (AF) cells from rabbit intervertebral disc. We seeded NP cells into the inner core of the hybrid scaffold and AF cells into the outer portion of it. Cellular viability and proliferation were assayed by 3-(4,5-dimethylthiazole-2-yl) -2,5- diphenyltetrazolium -bromide (MTT) test. PLGA and PLGA/DBP scaffolds were implanted in subcutaneous of athymic nude mouse to observe the formation of disc-like tissue in vivo. And then we observed change of morphology and hematoxylin and eosin (H&E). Formation of disc-like tissue was better DBP/PLGA hybrid scaffold than control. Specially, we confirmed that scaffold impregnated 20 and 40% DBP affected to proliferation of disc cell and formation of disc-like tissue.

• Macromolecular Research
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• v.10 no.3
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• pp.158-167
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• 2002

In order to endow with new bioactive functionality from small intestine submucosa (SIS) powder as natural source to poly (L-lactide) (PLA) and poly (lactide-co-glycolide) (PLGA) synthetic biodegradable polymer, porous SIS/PLA and SIS/PLGA as natural/synthetic composite scaffolds were prepared by means of the solvent casting/salt leaching methods for the possibility of the application of tissue engineered bone and cartilage. A uniform distribution of good interconnected pores from the surface to core region was observed the pore size of 40~500 ${\mu}{\textrm}{m}$ independent with SIS amount using the solvent casting/salt leaching method. Porosities, specific pore areas as well as pore size distribution also were almost same. After the fabrication of SIS/PLA hybrid scaffolds, the wetting properties was greatly enhanced resulting in more uniform cell seeding and distribution. Five groups as PGA non-woven mesh without glutaraldehyde (GA) treatment, PLA scaffold without or with GA treatment, and SIS/PLA (Code No.3 ; 1 : 12 of salt content, (0.4 : 1 of SIS content, and 144 ${\mu}{\textrm}{m}$ of median pore size) without or with GA treatment were implanted into the back of nude mouse to observe the effect of SIS on the induction of cells proliferation by hematoxylin and eosin, and von Kossa staining for 8 weeks. It was observed that the effect of SIS/PLA scaffolds with GA treatment on bone induction are stronger than PLA scaffolds, that is to say, in the order of PLA/SIS scaffolds with GA treatment > PLA/SIS scaffolds without GA treatment > PGA nonwoven > PLA scaffolds only with GA treatment = PLA scaffolds only without GA treatment for the osteoinduction activity. The possible explanations are (1) many kinds of secreted, circulating, and extracellular matrix-bound growth factors from SIS to significantly affect critical processes of tissue development and differentiation, (2) the exposure of SIS to GA resulted in significantly calcification, and (3) peri-implant fibrosis due to covalent bonding between collagen molecule by crosslinking reaction. In conclusion, it seems that SIS plays an important role for bone induction in SIS/PLA scaffolds for the application of tissue engineering area.

• Journal of Periodontal and Implant Science
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• v.38 no.sup2
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• pp.273-298
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• 2008

Purpose: The aim of this review is to introduce a novel bone-graft material for hard-tissue regeneration based on the calcium phosphate glass(CPG). Materials and Methods: CPG was synthesized by melting and subsequent quenching process in the system of CaO-$CaF_2-P_2O_5$-MgO-ZnO having a much lower Ca/P ratio than that of conventional calcium phosphates such as HA or TCP. The biodegradability and bioactivity were performed. Effects on the proliferation, calcification and mineralization of osteoblast-like cells were examined in vitro. Influence in new bone and cementum formations was investigated in vivo using calvarial defects of Sprague-Dawley rats as well as 1-wall intrabony defect of beagle dogs. The application to the tissue-engineered macroporous scaffold and in vitro and in vivo tests was explored. Results: The extent of dissolution decreased with increasing Ca/P ratio. Exposure to either simulated body fluid or fetal bovine serum caused precipitation on the surface. The calcification and mineralization of osteoblast-like cells were enhanced by CPG. CPG promoted new bone and cementum formation in the calvarial defect of Sprague-Dawley rats after 8 weeks. The macroporous scaffolds can be fabricated with $500{\sim}800{\mu}m$ of pore size and a three-dimensionally interconnected open pore system. The stem cells were seeded continuously proliferated in CPG scaffold. Extracellular matrix and the osteocalcin were observed at the $2^{nd}$ days and $4^{th}$ week. A significant difference in new bone and cementum formations was observed in vivo (p<0.05). Conclusion: The novel calcium phosphate glass may play an integral role as potential biomaterial for regeneration of new bone and cementum.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.36 no.4
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• pp.243-249
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• 2010

Tissue engineered bone (TEB) can replace an autogenous bone graft requiring an secondary operation site as well as avoid complications like inflammation or infection from xenogenic or synthetic bone graft. Adult mesenchymal stem cells (MSC) for TEB are considered to have various ranges of differentiation capacity or multipotency by the donor site and age. This study examined the effect of age on proliferation capacity, differentiation capacity and bone morphogenetic protein-2 (BMP-2) responsiveness of human bone marrow stromal cells (hBMSC) according to the age. In addition, to evaluate the effect on enhancement for osteoblast differentiation, the hBMSC were treated with Trichostatin A (TSA) and 5-Azacitidine (5-AZC) which was HDAC inhibitors and methyltransferase inhibitors respectively affecting chromatin remodeling temporarily and reversibly. The young and old group of hBMSC obtained from the iliac crest from total 9 healthy patients, showed similar proliferation capacity. Cell surface markers such as CD34, CD45, CD90 and CD105 showed uniform expression regardless of age. However, the young group showed more prominent transdifferentiation capacity with adipogenic differentiation. The osteoblast differentiation capacity or BMP responsiveness was low and similar between young and old group. TSA and 5-AZC showed potential for enhancing the BMP effect on osteoblast differentiation by increasing the expression level of osteogenic master gene, such as DLX5, ALP. More study will be needed to determine the positive effect of the reversible function of HDAC inhibitors or methyltransferase inhibitors on enhancing the low osteoblast differentiation capacity of hBMSC.