• Transactions of the Korean Society of Mechanical Engineers A
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• v.41 no.7
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• pp.645-653
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• 2017

In bone tissue engineering, polycaprolactone (PCL) is one of the most widely used biomaterials in the manufacturing of scaffolds as a synthetic polymer having biodegradability and biocompatibility. The strut width in the fabrication of scaffolds is an important part of tissue regeneration in in-vitro and in-vivo experiments, because it affects not only the pore size but also the porosity. In this study, we used polymer deposition system (PDS) and design of experiments (DOE) to explore the optimal process conditions to achieve a systematic and efficient scaffold manufacturing process, using temperature, pressure, scan velocity, and nozzle tip height as the parameters for the experiments. The aim of this research was to fabricate a 3D PCL scaffold having a uniform strut width of $150{\mu}m$ using DOE; it was proved that the strut width was constant in all the experimental groups by fabricating the PCL scaffolds according to various pore patterns as well as one pore pattern.

• Journal of Korean Neurosurgical Society
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• v.46 no.4
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• pp.397-402
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• 2009

Objective : In this study, the authors assessed the ability of rat bone marrow derived mesenchymal stem cells (BMDMSCs), in the presence of a growth factor, fibroblast growth factor-4 (FGF-4) and hydroxyapatite, to act as a scaffold for posterolateral spinal fusion in a rat model. Methods : Using a rat posterolateral spine fusion model. the experimental study comprised 3 groups. Group 1 was composed of 6 animals that were implanted with 0.08 gram hydroxyapatite only. Group 2 was composed of 6 animals that were implanted with 0.08 gram hydroxyapatite containing $1{\times}10^6/60{\mu}L$ rat of BMDMSCs. Group 3 was composed of 6 animals that were implanted with 0.08 gram hydroxyapatite containing $1{\times}10^6/60{\mu}L$ of rat BMDMSCs and FGF-4 $1{\mu}G$ to induce the bony differentiation of the BMDMSCs. Rats were assessed using radiographs obtained at 4, 6, and 8 weeks postoperatively. After sacrifice, spines were explanted and assessed by manual palpation, high-resolution microcomputerized tomography, and histological analysis. Results : Radiographic, high-resolution microcomputerized tomographic, and manual palpation revealed spinal fusion in five rats (83%) in Group 2 at 8 weeks. However, in Group 1, three (60%) rats developed fusion at L4-L5 by radiography and two (40%) by manual palpation in radiographic examination. In addition, in Group 3, bone fusion was observed in only 50% of rats by manual palpation and radiographic examination at this time. Conclusion : The present study demonstrates that 0.08 gram of hydroxyapatite with $1{\times}10^6/60{\mu}L$ rat of BMDMSCs induced bone fusion. FGF4, added to differentiate primitive $1{\times}10^6/60{\mu}L$ rat of BMDMSCs did not induce fusion. Based on histologic data, FGF-4 appears to induce fibrotic change rather than differentiation to bone by $1{\times}10^6/60{\mu}L$ rat of BMDMSCs.

• Macromolecular Research
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• v.9 no.5
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• pp.267-276
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• 2001

In order to endow with new bioactive functionality from demineralized bone particle (DBP) as natural source to poly(L-lactide) (PLA) synthetic biodegradable polymer, porous DBP/PLA as natural/synthetic composite scaffolds were prepared and compared by means of the emulsion freeze drying and solvent casting/salt leaching methods for the possibility of the application of tissue engineered bone and cartilage. For the emulsion freeze drying method, it was observed that the pore size decreased in the order of 79$\mu\textrm{m}$ (PLA control) > 47$\mu\textrm{m}$ (20% of DBP) > 23 $\mu\textrm{m}$ (40% of DBP) > 15$\mu\textrm{m}$ (80% of DBP). Porosities as well as specific pore areas decreased with increasing the amount of DBR. It can be explained that DBP acts like emulsifier resulting in stabilizing water droplet in emulsion. For the solvent casting/salt leaching method, a uniform distribution of well interconnected pores from the surface to core region were observed the pore size of 80 ∼70 $\mu\textrm{m}$ independent with DBP amount. Porosities as well as specific pore areas also were almost same. For pore size distribution by the mercury intrusion porosimeter analysis between the two methods, the pore size distribution of the emulsion freeze drying method was broader than that of the solvent casting/salt leaching method due to the mechanism of emulsion formation. Scaffolds of PLA alone, DBP/PLA of 40 and 80%, and DBP powder were implanted on the back of athymic nude mouse to observe the effect of DBP on the induction of cells proliferation by hematoxylin and eosin staining for 8 weeks. It was observed that the effect of DBP/PLA scaffolds on bone induction are stronger than PLA scaffolds, even though the bone induction effect of DBP/PLA scaffold might be lowered than only DBP powder, that is to say, in the order of DBP only > DBP/PLA scaffolds of 40 and 80% DBP > PLA scaffolds only for osteoinduction activity. In conclusion, it seems that DBP plays an important role for bone induction in DBP/PLA scaffolds for the application of tissue engineering area.

• Archives of Plastic Surgery
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• v.38 no.2
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• pp.155-160
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• 2011

Purpose: Fibrillar collagens like type I collagen, are the major constituent of the extracellular matrix and structural protein of bone. Also, it can be a scaffold for osteoblast migration. The purpose of this study is to estimate the effects of absorbable atelo-collagen sponge (Teruplug$^{(R)}$, Terumo biomaterials Co., Tokyo, Japan) insertion in tooth extraction sites on periodontal healing of the second molar, healing of the fractured mandibular bone and new bone formation of third molar socket after the extraction of the impacted third molar with mandibular angle fracture. Methods: In our study of six cases of mandibular angle fractures, all of them underwent the extraction of the third molar tooth & absorbable atelo-collagen sponge insertion in tooth extraction site. Three of them had a intraoral infection & oral opening to fracture site, two of the six had dental caries, and only one had reduction problem due to third molar position. Six consecutive patients with noncomminuted fractures of the mandibular angle were treated by open reduction and internal fixation using one noncompression miniplates and screws placed through a transoral incision. Results: All of the patients have showed good postoperative functions and have not experienced complications requiring second surgical intervention. There was well healing of the mandibular bone and the most new bone formation of third molar socket after the extraction of the impacted third molar with mandibular angle fracture. Conclusion: The results of this study suggest that absorbable atelo-collagen sponge is relatively favorable bone void filler with prevention of tissue collapse, food packing, and enhance periodontal healing. Thus, the use of atelo-collagen sponge and one noncompression miniplate seems to be relatively easy, safe, and effective for the treatment of fractures of the mandibular angle and third molar extraction.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.36 no.5
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• pp.366-374
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• 2010

Introduction: This study evaluated the capability of silk fibroin (SF) and recombinant human bone morphogenetic protein-2 loaded SF (SF-BMP) as a bone defect replacement matrix when grafted in a calvarial bone defect of rats in vivo. Materials and Methods: A total 70 calvarial critical size defects (5.0 mm in diameter) made on 35 adult female Sprague-Dawley rats were used in this study. The defects were transplanted with (1) rhBMP-2 loaded silk fibroin graft (SF-BMP: 0.8+$10\;{\mu}g$), (2) Silk fibroin (SF: $10\;{\mu}g$), and (3) no graft material (Raw). The samples were evaluated with soft x-rays, alkaline phosphatase activity, calcium/phosphate quantification, histological and histomorphometric analysis at postoperative 4 and 8 weeks. Results: The SF-BMP group ($48.86{\pm}14.92%$) had a significantly higher mean percentage bone area than the SF group ($24.96{\pm}11.01%$) at postoperative 4 weeks.(P<0.05) In addition, the SF-BMP group ($40.01{\pm}12.43%$) had a higher % bone area at postoperative 8 weeks than the SF group ($33.26{\pm}5.15%$). The mean ratio of gray scale levels to the host bone showed that the SF-BMP group ($0.67{\pm}0.08$) had a higher mean ratio level than the SF group ($0.61{\pm}0.09$) at postoperative 8 weeks. These differences were not statistically significant.(P=0.168 and P=0.243, respectively) The ratio of the calcium and phosphate contents of the SF-BMP ($0.93{\pm}0.22$) group was lower than that of the SF ($1.90{\pm}1.42$) group at postoperative 4 weeks. However, the SF-BMP group ($0.75{\pm}0.31$) had a higher Ca/$PO_4$ ratio than the SF ($0.68{\pm}0.04$) at postoperative 8 weeks. These differences were not statistically significant.(P=0.126 and P=0.627, respectively) For the bone-specific alkaline phosphatase (ALP) activity, which is recognized as a reliable indicator of the osteoblast function, the SF-BMP ($23.71{\pm}8.60\;U/L$) groups had a significantly higher value than the SF group ($12.65{\pm}6.47\;U/L$) at postoperative 4 weeks.(P<0.05) At postoperative 8 weeks, the SF-BMP ($21.65{\pm}10.02\;U/L$) group had a lower bone-specific ALP activity than the SF group ($16.72{\pm}7.35\;U/L$). This difference was not statistically significant.(P=0.263) For the histological evaluation, the SF-BMP group revealed less inflammation, lower foreign body reactions and higher bone healing than the SF group at postoperative 4 and 8 weeks. The SF group revealed more foreign body reactions at postoperative 4 weeks. However, this immunogenic reaction decreased and the remnant of grafted material was observed at postoperative 8 weeks. For histomorphometric analysis, the SF-BMP group had a significantly longer bone length to total length ratio than those of the SF group at postoperative 4 and 8 weeks.(P<0.05) Conclusion: The rhBMP-2 loaded silk fibroin graft revealed fewer immunoreactions and inflammation as well as more new bone formation than the pure silk fibroin graft. Therefore, silk fibroin may be a candidate scaffold for tissue engineered bone regeneration.

• 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.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.38
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• pp.2.1-2.9
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• 2016

Background: We reviewed the biological and mechanical properties of porous hydroxyapatite (HA) compared to other synthetic materials. Computer-aided design/computer-aided manufacturing (CAD/CAM) was also evaluated to estimate its efficacy with clinical and radiological assessments. Method: A systematic search of the electronic literature database of the National Library of Medicine (PubMed-MEDLINE) was performed for articles published in English between January 1985 and September 2013. The inclusion criteria were (1) histological evaluation of the biocompatibility and osteoconductivity of porous HA in vivo and in vitro, (2) evaluation of the mechanical properties of HA in relation to its porosity, (3) comparison of the biological and mechanical properties between several biomaterials, and (4) clinical and radiological evaluation of the precision of CAD/CAM techniques. Results: HA had excellent osteoconductivity and biocompatibility in vitro and in vivo compared to other biomaterials. HA grafts are suitable for milling and finishing, depending on the design. In computed tomography, porous HA is a more resorbable and more osteoconductive material than dense HA; however, its strength decreases exponentially with an increase in porosity. Conclusions: Mechanical tests showed that HA scaffolds with pore diameters ranging from 400 to $1200{\mu}m$ had compressive moduli and strength within the range of the human craniofacial trabecular bone. In conclusion, using CAD/CAM techniques for preparing HA scaffolds may increase graft stability and reduce surgical operating time.

• BMB Reports
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• v.43 no.1
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• pp.52-56
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• 2010

Silk fibroin, produced by the silkworm Bombyx mori, has been widely studied as a scaffold in tissue engineering. Although it has been shown to be slowly biodegradable, cellular responses to degraded silk fibroin fragments are largely unknown. In this study, silk fibroin was added to MG-63 cell cultures, and changes in gene expression in the MG-63 cells were screened by DNA microarray analysis. Genes showing a significant (2-fold) change were selected and their expression changes confirmed by quantitative RT-PCR and western blotting. DNA microarray results showed that alkaline phosphatase (ALP), collagen type-I alpha-1, fibronectin, and transforming growth factor-${\beta}1$ expressions significantly increased. The effect of degraded silk fibroin on osteoblastogenic gene expression was confirmed by observing up-regulation of ALP activity in MG-63 cells. The finding that small fragments of silk fibroin are able to increase the expression of osteoblastogenic genes suggests that controlled degradation of silk fibroin might accelerate new bone formation.

• Macromolecular Research
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• v.13 no.4
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• pp.285-292
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• 2005

We developed alginate beads loaded with transforming growth $factor-{\beta}_{1}(TGF-{\beta}_{1})$ to examine the possible application of the scaffold and cytokine carrier in tissue engineering. In this study, bone marrow stromal cells (BMSCs) and $TGF{\beta}_{1}$ were uniformly encapsulated in the alginate beads and then cultured in vitro. The cell morphology and shape of the alginate beads were observed using inverted microscope, scanning electron microscope (SEM), histological staining and RT-PCR to confirm chondrogenic differentiation. The amount of the $TGF{\beta}_{1}$ released from the $TGF-{\beta}_{1}$ loaded alginate beads was analyzed for 28 days in vitro in a phosphate buffered saline (pH 7.4) at $37^{\circ}C$. We observed the release profile of $TGF-{\beta}_{1}$ from $TGF-{\beta}_{1}$ loaded alginate beads with a sustained release pattern for 35 days. Microscopic observation showed the open cell pore structure and abundant cells with a round morphology in the alginate beads. In addition, histology and RT-PCR results revealed the evidence of chondrogenic differentiation in the beads. In conclusion, these results confirmed that $TGF-{\beta}_{1}$ loaded alginate beads provide excellent conditions for chondrogenic differentiation.

• Korean Journal of Veterinary Research
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• v.58 no.4
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• pp.211-217
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• 2018

Mesenchymal stem cells (MSCs) are useful candidates for tissue engineering and cell therapy. Physiological cell environment not only connects cells to each other, but also connects cells to the extracellular matrix that provide mechanical support, thus exposing the entire cell surface and activating signaling pathways. Hydrogel is a polymeric material that swells in water and maintains a distinct 3-dimensional (3D) network structure by cross linking. In this study, we investigated the optimized cellular function for canine adipose tissue-derived MSCs (cAD-MSCs) using hydrogel. We observed that the expression levels of Ki67 and proliferating cell nuclear antigen, which are involved in cell proliferation and stemness, were increased in transwell-hydrogel (3D-TN) compared to the transwell-normal (TN). Also, transforming growth factor-${\beta}1$ and SOX9, which are typical bone morphogenesis-inducing factors, were increased in 3D-TN compared to the TN. Collagen type II alpha 1, which is a chondrocyte-specific marker, was increased in 3D-TN compared to the TN. Osteocalcin, which is a osteocyte-specific marker, was increased in 3D-TN compared to the TN. Collectively, preconditioning cAD-MSCs via 3D culture systems can enhance inherent secretory properties that may improve the potency and efficacy of MSCs-based therapies for bone regeneration process.