Papers by Author: Hong In Shin

Abstract: To improve the efficiency of osteogenic repair, we compared 3 types of round granular bone substitutes composed of hydroxyapatite (HA) in a single opened large pore covered by one or more thin shell structure, biphasic HA and tricalcium phosphate (TCP) in a compact granules with small uniform interconnected internal pores, and bioglass(BG) in a compact granules with hierarchical interconnected pores its bone repair efficiency by evaluation of cellular toxicity, cellular attachment and proliferation rate, and osteogenic supportive effect. They were nontoxic and revealed no noxious effect on cellular proliferation and osteoblastic differentiation. The cultured cells were most effectively proliferated on HA granular bone substitute surface. However, the bony repair of calvarial defects was most effective by BCP granular bone substitutes. The implanted BCP and HA granular bone substitutes showed excellent osteoconductive bone growth and favorable bone regeneration within 3 weeks compared to BG granular bone substitutes. All type granular bone substitutes were well incorporated into newly formed bone without foreign body reaction. Except for HA granular bone substitute, some implanted BG and BCP granular bone substitutes were partially resobed by TRAP positive multinucletated cells. These findings suggest that round granular biphasic calcium phosphate bone substitute structured with fully interconnected uniform sized internal pore might be a more promising bone substitute for small-sized none load-bearing bone defects.

Abstract: To develop a suitable scaffold for tissue-engineered bone regeneration, we compared the efficiency of tissue-engineered bone regeneration according to the porous structure of calcium metaphosphate (CMP) ceramic scaffolds. Each scaffold was prepared with a sponge method and a foam-gel method, respectively. Both scaffolds, having either interconnected trabecular pores formed by the sponge method or fully interconnected globular pores formed by the foam-based technology, were not cytotoxic and elicited neither an immune nor an inflammatory response regardless of geometry and fabrication method. The fully interconnected globular porous scaffold showed more favorable compression strength and facilitated osteogenic repair by favoring cellular attachment and osteogenic differentiation with good osteoconductivity compared to the interconnected trabecular pore structured scaffold. These results suggest that the fully interconnected globular porous structure would be more suitable for both a bone substitute and scaffold for bioactive material-based or cell-based tissue bone regeneration.

Abstract: Recently, nanomaterials have received considerable attention because of their potential applications in the biomedical field. In the present study, we investigated the effects of nano-sized calcium metaphosphate (CMP) particles (50 nm) compared with micro-sized CMP particles (200-500 nm and 10 μm) on the proliferation and osteoblastic differentiation of human bone marrow stem cells (BMSCs). BMSCs were challenged with CMP particles with different sizes for 3, 5, and 7 days. An analysis of the proliferation revealed that the nano-sized CMP particles (50 nm) stimulated the proliferation of BMSCs up to 27.79% compared to the untreated control. This stimulatory effect of the nano-sized CMP particle was dose-dependent. CMP particles appeared to adhere on the surface of BMSCs but this did not cause distinguishable morphological changes. Moreover, all CMP particles (50 nm to 10 μm) were capable of stimulating an osteoblastic differentiation of BMSCs as accessed by alkaline phosphatase (ALP) and von Kossa stainings. Further molecular analysis revealed that all the CMP particles induced an expression of osteoblast-related genes such as osteocalcin (OC) and collagen I (Col I). Taken together, our data demonstrate that nano-sized CMP particles have the potential to stimulate the proliferation and osteoblastic differentiation of BMSCs.

Abstract: An appropriate scaffold, which provides structural support for transplanted cells and acts as a vehicle for the delivery of biologically active molecules, is critical for tissue engineering. We developed a fully interconnected globular porous biphasic calcium phosphate ceramic scaffold by adopting a foaming method, and evaluated its efficiency as a bone substitute and a scaffold for bone tissue engineering by in vitro and in vivo biocompatible analysis and its osteogenic healing capacity in rat tibial bone defects. They have spherical pores averaging 400um in diameter and interconnecting interpores averaging 70um in diameter with average 85% porosity. They elicited no cytotoxicity and noxious effect on cellular proliferation and osteoblastic differentiation during the cell-scaffold construct formation. Also the bone defects grafted with fully interconnected globular porous biphasic calcium phosphate ceramic blocks revealed excellent bone healing within 3 weeks. These findings suggest that the fully interconnected porous biphasic calcium phosphate scaffold formed by the foaming method can be a promising bone substitute and a scaffold for bone tissue engineering.

Abstract: To develop a suitable scaffold optimizing bone regeneration, we developed bovine bone ash derived fully connected porous HA ceramic scaffolds adopting a foaming method. They revealed excellent biocompatibility. The attached cells on the scaffolds proliferated in multi-layers with osteoblastic differentiation. The bone defects grafted with bovine bone ash derived fully interconnected porous HA ceramics having average 500 μm sized spherical pores and average 150 μm sized interconnecting interpores with average 80% porosity were favorably healed without any pathologic changes within 3 weeks. New bone ingrowth with excellent osteoconduction through the spherical pores along the inner surface was noted from 1 week after implantation. Each spherical pore was filled with hematopoietic marrow and newly formed bone which with time was well integrated with the porous HA ceramic scaffold with time. These findings suggest that the bovine bone ash-derived fully interconnected porous HA ceramic formed by foaming method can be a promising bone substitute and a scaffold for bone tissue engineering.

Abstract: To improve ostegenic healing efficiency by demineralized bone matrix, we evaluated the ectopic bone formation induced by variously demineralized allogenic cortical bone matrices at subcutaneous and muscular sites in rats. The rat tubular cortical bone matrices were demineralized in heated 0.6N HCl at 60 °C for 5 and 20 mins, respectively, using a controlledheat ultrasonic cleaner and implanted in rat dorsal subcutaneous pouches and thigh muscles for 1-3 weeks. The influence of the demineralized condition of bone matrix on cellular proliferation and osteogenic differentiation was also evaluated in vitro by MTT assay and ALP staining. The cortical matrices were completely demineralized within 20 mins by sonication and heating of diluted 0.6 N HCl. The sonicated bone matrices in heated acidic solution at 60 °C revealed no adverse immunogenic and inflammatory response in vivo regardless of demineralized condition. Cellular proliferation and osteoblastic differentiation was facilitated by more fully demineralized. Ectopic bone formation was induced only by demineralized bone matrices and were more favorable in fully demineralized matrices. The ectopic bone induction was more favorably in subcutaneous pouches than in muscular tissue. These findings suggest that a fully demineralized cortical bone matrix maximizes osteogenic repair by exposing more bioactive molecules which in turn induce chondro- and osteognic differentiation of mesenchymal cells around the implanted matrices, and that the sonication of diluted 0.6 N HCl heated at 60 ° C is a rapid and effective method for sterile demineralized graft preparation.

Abstract: Despite many outstanding research works on cartilage tissue engineering, actual clinical application is not quite successful because of the absorption and progressive distortion of tissue engineered cartilage. We have developed a new method of cartilage tissue engineering comprising chondrocyte mixed Pluronic F-127 and cultured chondrocyte cell sheet which entirely cover the cell-Pluronic complex. We believe the addition of cultured chondrocyte cell sheet enhances the efficacy of chondrogenesis in vivo. Human ear cartilage piece was enzymatically dissociated and chondrocyte suspension was acquired. Chondrocytes were cultured and expanded as the routine manner. Cultured chondrocytes were plated in high-density monolayer and cultured with Chondrogenic media in 5% CO2 incubator. After 3 weeks of culture, chondrocyte cell sheet was formed and complete single sheet of chondrocyte could be harvested by gentle manipulation of culture plate with a cell scraper. Chondrocyte-Pluronic complex was established by mixing 1x 106 cells with 0.5 of Pluronic F- 127. Chondrocyte-Pluronic complex was completely covered with a sheet of cultured chondrocyte. The completed tissue engineered constructs were implanted into the subcutaneous tissue pocket of nude mice on the back. Tissue engineered constructs without cultured cell sheet were used as control. Samples were harvested at 8 weeks postoperatively and they were subjected to histological analysis and assayed for glycosaminoglycan (GAG), and type II collagen. Grossly, the size of cartilage specimen of cultured chondrocyte cell sheet covered group was larger than that of the control. On histologic examination, the specimen of cultured chondrocyte cell sheet covered group showed lacunae-containing cells embedded in a basophilic matrix. The chondrocyte cell sheet covered group specimen resembled mature or immature cartilage. The result of measurement of GAG and type II collagen of cartilage specimen of cultured chondrocyte sheet covered group was higher than that of the control. In conclusion, the new method of cartilage tissue engineering using chondrocyte cell sheet seems to be an effective method providing higher cartilage tissue gain and reliable success rate for cartilage tissue engineering.

Abstract: As a part of the effort to develop a suitable scaffold for tissue-engineered bone regeneration, we modified calcium metaphosphate (CMP) ceramic with Na20 and evaluated its efficiency as a scaffold. We incorporate 5% Na20 into pure CMP and prepare for an average pore size of 250 or 450 µm average pore sizes. The incorporation of 5% Na2O caused reduced compressive strength and there was no change in biodegradability. The in vitro cellular attachment and proliferation rate, however, were slightly improved. The 5% Na2O-incorporated macroporous CMP ceramic-cell constructs treated with Emdogain induced ectopic bone formation more effectively than those without Emdogain treatment. These results suggest that the incorporation of 5% Na2O into pure CMP is not effective for improving the physical characteristics of pure CMP but it is positive for improving the cellular reaction and osteogenic effect with the addition of Emdogain.

Abstract: Five kinds of gypsums, (1) CaSO4•2H2O (caldium sulfate dihydrate; CSD), (2) CaSO4•1/2H2O (calcium sulfate hemihydrate; CSH), (3) CaSO4 (calcium sulfate anhydrite; CSA), (4) CSH200 (CSH heat-treated at 200°C after self-hardening), and (5) CSH600 (CSH heat-treated at 600°C after self-hardening) were used as candidates for coating materials on calcium metaphosphate (CMP) scaffod to control degradation rate of CMP and to extend degradation limit. The disks of CSD, CSH, CSA, CSH 200, and CSH600 were prepared by self-hardening after mixing with water, where CSH200 and CSH600 were heat-treated at 200°C and 600°C, respectively. In order to control fast resorption rate of gypsum, CMP-CSA composites were prepared with different CSA contents such as 0, 5, 10, 20, 30, 50, and 70 vol% and heat-treated at 900°C for 4 hours. The degradation rates of various gypsums were evaluated in revised simulated body fluid (r-SBF) for 1, 3, 7, and 21 days, respectively. Degradation rate of each specimen was measured in terms of weight loss change with time and degraded surface morphology was examined by SEM. All kinds of gypsums were transformd into CSD after self-hardening with water. Most of gypsums were degraded by 35~60 wt% at 7 days and by 70~99 wt% at 21 days of soaking in SBF. In the group of CMP-CSA composites, the degree of degradation of them was considerably retarded compared to that of five pure gypsums. The surface morphology showed elongated needle-like crystals during the degradation with time.

Abstract: For effective bone regeneration, various surface modifications have been tried. In an effort to improve osteogenic repair potential, we evaluated recombinant peptides containing the RGD domain as a bioactive molecule for tissue-engineered bone regeneration. The synthetic peptides slightly suppressed cellular proliferation in the in vitro culture system but induced favorable osteoblastic differentiation, which was determined by MTT and ALP activity staining, respectively. The synthetic peptide coated CMP granules, which were implanted into the mandibular bone defects showed more favorable bone repair compared to the non-coated CMP implantation. In addition, there were not any sign of inflammatory reaction. These findings suggest that synthesized peptides containing the RGD domain enhance cellular attachment and osteogenic activity in vivo condition and that the peptide-coated CMP granules can serve as a biocompatible bone substitute.