초록 ▼

○ 본 연구는 세계 최초로 시도되는 분야로서, 방사선을 이용하여 약물 구강 점막 부착층과 약물 보호층 이중구조로 구성된 구강점막 부착용 약물전달체로서, 독성이 없고 점막접착성이 우수하며, 서방성 약물 방출 특성을 가지고 있어 구내염 치료에 탁월한 효과를 나타냄.
○ 총 8종 이상의 개질된 고분자 하이드로겔을 개발하였으며, 80% 이상의 높은 겔화율과 60 kPa 이상의 우수한 접착력을 나타냈으며, 4종류의 균주에 대하여 우수한 항균성을 나타냄.
○ 48 시간 까지 지속된 약물방출 거동과 80% 이상 염증이 저감되는 동물

○ 본 연구는 세계 최초로 시도되는 분야로서, 방사선을 이용하여 약물 구강 점막 부착층과 약물 보호층 이중구조로 구성된 구강점막 부착용 약물전달체로서, 독성이 없고 점막접착성이 우수하며, 서방성 약물 방출 특성을 가지고 있어 구내염 치료에 탁월한 효과를 나타냄.
○ 총 8종 이상의 개질된 고분자 하이드로겔을 개발하였으며, 80% 이상의 높은 겔화율과 60 kPa 이상의 우수한 접착력을 나타냈으며, 4종류의 균주에 대하여 우수한 항균성을 나타냄.
○ 48 시간 까지 지속된 약물방출 거동과 80% 이상 염증이 저감되는 동물 실험 결과를 나타냄.
○ 본 연구는 세계 최초로 방사선 기술을 이용하여 천연고분자 및 천연/합성고분자 복합체기반의 지지체의 개발은 물론 동물실험 대체용 인공피부 제조기술을 확립하였음. 특히,개발된 인공피부용 지지체의 표면 특성 개선기술과 세포 흡착 및 증식능을 조절할 수 있는 자체기술을 개발하여 다양한 기능기들의 도입 및 도입량 조절이 가능해짐.
○ 총 8종의 인공피부용 고분자 지지체를 개발하였으며, 80% 이상의 높은 겔화율과 기계적물성, 80% 이상의 생체적합성 및 150% 이상의 세포 흡착율과 세포 증식능을 가지는 지지체를 개발하였음.

( 출처 : 요약서 6p )

Abstract ▼

Ⅳ. The result of the research and development
■ Development of hydrogel for buccal mucosa adehsion
○ Development of PAA hydrogel having duplex configuration
- The gel fraction increased with the increase in the content of PAA and irradiatiom dose. The mucoadhesive strength was evaluated by

Ⅳ. The result of the research and development
■ Development of hydrogel for buccal mucosa adehsion
○ Development of PAA hydrogel having duplex configuration
- The gel fraction increased with the increase in the content of PAA and irradiatiom dose. The mucoadhesive strength was evaluated by measuring the force required to detach the formulation from oral mucosa tissue of pigs using a universal test machine. As the increased irradiation dose, the mucoadhesive force was increased.
○ Development of PAA/PEG hydrogel for buccal mucosa adhesion
- The gel fraction increased as annealing time increased. The gel strength decreased with an increase in PEG content. The compressive strength of the hydrogels was increased with an increase in annealing time and with a decrease PEG concentration. The probable reason of this result is that the presence of PEG normally increases the elasticity of the gel due to the plasticizing effect. As the contents of PEG increased from 0.25 to 0.5%, the mucoadhesive force was increased, except for the formulation with 1%.
○ Development of PV A/PEG hydrogel for buccal mucosa adhesion
- The effects of PEG content and annealing of PVA hydrogels were obsered. The gel fraction increased as the content of PEG decreased and annealing time increased. The degree of swelling decreased with increasing PEG content and decreasing annealing time. Also, the thermal and mechanical properties increased with the PEG concentration and annealing time.
○ Development of micro-hydrogel
- The PAA micro- hydrogel have nearly a perfect sphere. Also, the average particle size decreased with an increasing irradiation dose due to the intra-and inter-crosslinking. As the electron beam irradiation dose increased, the absolute values of zeta potential increased.

■ Development of hydrogel containing drug for buccal mucosa adehsion
○ Effect and safety evaluation of PAA hydrogel containing drug
- The PAA hydrogel having duplex configuration have been investigated for inhibiting the growth of S. aureus and E. coli, S. epidermidis, C. albican on solid growth media. The antibacterial tests indicated that the hydrogels had good antibacterial activities.
- Triamcinolone acetonide was released constantly from the gel formulations at 37℃ and could reach 100% at about 6 hour.
- The cytotoxicity of the PAA hydrogel were evaluated by using CCK-8 assay.The PAA hydrogels were considered not to be cytotoxic.
- The PAA hydrogels displayed the eminent healing result from the animal tests and clinical trials.
○ Development of PVA/PEG hydrogel containing AgNPs
- The characteristics of Ag NPs in the PVA/PEG hydrogels were measured by UV-vis, FE SEM, Particle size analyzer. The UV-vis analysis indicated that the concentration of Ag NPs was enhanced by increasing of radiation dose from 1 to 10 kGy. Ag NPs had smaller sizes and shapes with an increasing radiation dose.
○ Development of CMC hydrogel contaimng AgNPs
- The intensity of the absorption band of AgNPs increases sigmficantly and shift to the blue (from 440 to 420 nm) with increasing the irradiation dose up to 5 kGy. The absorption spectra became much narrower, and intense UV spectra of the samples shows that smaller-diameter Ag NPs were obtained at larger radiation doses. Ag NPs had smaller sizes and shapes with an increasing radiation dose. The CMC hydrogel contaimng AgNPs have been investigated for inhibiting the growth of Staphylococcus aureus and Escherichia coli strains in liquid as well as on solid growth media. The antibacterial tests indicated that the hydrogels contaimng Ag NPs have a good antibacterial activity.
○ Development of PAA hydrogel contaimng zinc oxide or zinc chloride
- Zinc chloride or zinc oxide based hydrogels have an effect on inhibiting and killing gram-positive S.aureus bacteria and gram-negative E.coli bacteria. Also the hydrogels contaimng a higher zinc concentration exhibit a much stronger inhibition effect, which confirms that the antibacterial activity is from the zinc partic1es not the PAAc composition.
○ Development of pluromc F127 hydrogel containing drug for buccal mucosa adhesion
- The thermosensitive and mucoadhesive gel formulations were prepared for buccal drug delivery using PF127 and carbopol. The naproxen-loaded gel showed a sol-gel transition around body temperature. The gelation temperature of the formulations was decreased, and mucoadhesive force, defined by detachment stress, was increased according to the concentration of carbopol. In vitro release was sustained with the addition of carbopol. Cross-linking the polymer gel by irradiation showed increased mechamcal strength and mucoadhesive force without major changes in in vitro release properties or cell viability.
- A naproxen-loaded thermoreversible gels showed a good sol-gel transition property and mucoadhesiveness could have potential for development as a more convement and effective means of treating oral disease with low toxicity.
- From the animal tests, the PAA hydrogels were proved to have the eminent healing effect.

■ Development of three dimensional (3D) scaffolds for cell culture by radiation
○ Fabrication of 3D porous scaffolds by radiation crosslinking/grafting
- 3D Porous natural polymer-based water-soluble chitosan/alginate/gelatin (WAG) scaffold was developed by using radiation crosslinking/grafting techniques instead of chemical reagent. The WAG scaffold has excellent mechanical strength by having amide/ester bond, moisture stability in phosphate buffer solution (37℃ , for 5 days), cell adhesion (> 80%) and compressive strength (> 200 kPa).
- In addition, 3D porous naturaνsynthetic polymer-based scaffold was developed by using radiation crosslinking/ grafting techniques. The structures of hyaluronic acid/ chondroitin sulfate/PVA (HA/CS/PVA) and hyaluronic acid/chondroitin sulfate/PAAc (HA/CS/PAAc) scaffolds were confirmed through FT-IR and NMR. These scaffolds have excellent gel fraction rate of over 70% and water content of more than 80%. In addition， biodegradation rate index was established by controlling the enzyme concentration and composition onto the scaffolds.
○ Fabrication of 3D porous scaffolds composed of biodegradable natural polymer by electrospinning method
- 3D Porous natural/biodegradable polymer-based nanofibers poly(L-lactide-co-caprolactone/marine collagen (PLCL/MC), gelatin/acrylic acid/PLCL (Gelatin/AAc/PLCL), RGD/acrylic acid/poly (L-lactic acid)(RGD/ AAc/PLLA) were developed by electrospinning having excellent tensile strength of 4.5 MPa or more, more than 80% of cell adhesion/ growth rate and racliation stability until 50 kGy of racliation dose.

■ Development of artificial skin model
○ Preparation of biocompatible film type scaffolds by racliation grafting
- Biocompatible film type scaffolds with average fiber diameter 700nm were developed with poly(L-lactide) (PLLA) , poly(L-lactide-co-ɛ
- caprolactone) (PLCL) using electrospinning technique and radiation technology.
- Optimal surface modification conditions using gamma-ray or electron-beam irradiation technology were achieved. Approximately 80% of AAc grafting yield was obtained under 15 kGy of gamma- ray exposure, and adsorption of TBO to AAc- grafted mesh prepared by electron-beam (20 kGy) was 400 μmol/mg.
- Especially, as a alternative technology for enhancing surface property of the scaffolds, carboxylic acid group and amine groups were introduced and controlled their introduction yield, and further introduction of hydroxyl group and amine groups were modulated exhibiting approimately 30% of introduction yield.
○ Development of cell culture technique for biodegradable scaffolds using adult stem cells
- Altemative control technology was developed for regulating surface hydrophilicity which can affect cell adhesion and proliferation. The results of cell adhesion analysis and proliferation of the cells on the hydrophilic scaffolds prepared by radiation technology exhibited that adhesion property of the cell on the mesh was increased to 150%, and long-term proliferation of the adherent cells was also achieved to 150% enhanced proliferation for 7 days of culture.
○ Development of artificia1 skin model
- The enzymatic degradation kinetics of polymeric artificial skin, HA/CS/PVA and HA/CS/PAAc scaffolds were established by controlling of their influence factors such as crosslinking density and composition of polymer matrix. When the drug molecular weight was less than 300(theophylline), the drug release rate was more than 95%, and the drug molecular weight was more than 300 (cefazoline), the drug release rate was more than 80%. In the solute permeation experimental results, when the drug molecular weight was less than 300 (theophylline), the drug permeation rate was more than 85%, and the drug molecular weight was more than 300 (cefazoline), the drug release rate was more than 60%.

■ Characterization and evaluation of artificia1 skin model
○ Optimization of ideal radiation doses for development of naturaνsynthetic polymer- based multi- functiona1 scaffolds
- Functionalization of electrospun nanofibrous meshes using radiation- based grafting technology
- After optimizing eletrospining condition of PLCL nanofibrous meshes(average fiber diameter: 700 nm), acrylic acid (AAc) grafted PLCL fibrous meshes were developed by optimizing gamma - ray irradiation doses and AAc concentration (AAc grafting amounts: 0.1- 1.2 mM/mg nanofiber).
- After EGF immobilization, which is essential bioactive molecules regulating cell proliferation and skin differentiation, introduction of EGF was analyzed by surface analysis and immobilization yield of EGF was calculated by ELISA. The immobilized amounts of EGF on the scaffolds were 8, 78, and 175 ng/mesh as 10, 100 and 200 ng/mL EGF solution reacted, the immobilization yield in the all experiments was higher and reached approximately 80%.
- Immobilization of bioactive molecules and optimization of immobilizing yield of bioactive molecules
○ Evaluation of cellular activity on the artificial skin
- The adherent profile of the hMSC on the scaffolds was investigated by analyzing fluorescent images. After 7 days of incubation, aspect ratio of adherent hMSC was increased to 2.5, 2.7, and 2.6 as compared with initia1 aspect ratio at day 1 (E-AP 10 ng: 1.12, 100 ng: 1.27, 200 ng: 1.33).
- In addition, the proliferation of hMSC on the scaffolds with different EGF amounts (10 to 200 ng/mL) was approximately 1.1- times increased, consequently, E-AP _200ng scaffolds successfully regulated the proliferation of hMSC for 7 days of culture exhibiting approximately 120% of enhanced proliferation.
- Development of co-immobilization technology of the bioactive molecules and ana1ysis of adhesion and proliferation of the hMSC on the multi-functional scaffolds
- As a additiona1 bioactive molecule to regulate cellulare behavior on the scaffolds, gelatin was further immobilized, approximately 200ug/mesh of gelatin was immobilized without overlapping EGF (EG- AP). In addition, after hMSC seeding and incubation on the EG-AP scaffolds, adhesion of hMSC was 1.4-times increased than those on the PLCL scaffolds at day 1. Those differences were increased at day 3 and 7, which were about 1.7-times and 2.4-times greater, respectively. The enhanc.ed proliferation of the cells on the EG-AP scaffolds also observed under fluorescent microscopy exhibiting that all of the cells cultured on the EG-AP scaffolds for 7 days completely covered the scaffolds without empty space.
○ Skin development of hMSC on the EG-AP scaffolds
- To prove skin differentiation of hMSC, involucrin, as a representative marker for skin differentiation, were stained after 14 days of incubation. The expression of involucrin on the scaffolds was approximately 2-3 times increased than those on the AP scaffolds exhibiting that skin differentiation may be facilitated by immobilized EGF.
○ Evaluation of testing methods of cosmetics.
- To evaluate toxicity of raw materials for cosmetics, representative antimicrobial agents were treated to hMSC-cultured EG-AP scaffolds. A variety of concentration of salicylic acid, formaldehyde, and methyl-paraben were used, all of sample showed reduced cell viability corresponding to each concentrations indicating that our novel artifical skin was acceptable to evaluate toxicity of raw materials for cosmetics.
- For the evaluation of skin corrosion of raw materials for cosmetics, different concentration of potassium hydroxide as a major corrosive materials were treated to hMSC- lived scaffolds, cell viability was decreased down to under 50% after 3 minutes and 19% after 60 minutes of treatments, which proved that novel artifical skin was useful to evaluate skin corrosion.
- Investigation of wrinkle- care materials effect on the artificial skin
- Investigation of wrinkle- care materials effect on the artificial skin was performed by measuring newly synthesized collagen. After 24 hours of wrinkle- care materials treatment (EGF solution), collagen expression or secretion was measured by ELISA. As 10 and 100 ng/mL of EGF were further treated, collagen expression was increased up to 130 and 140%, respectively. Additionally, when only 1ng/mL of EGF was treated the expression of collagen was 1.2-times greater than those of non-treated groups. Therefore, our artifical skin seems to be proper to evaluate wrinkle- care materials effect as like animal model.
- In the investigation of whitening materials effect on the artificial skin,EG-AP scaffolds did not regulate melanogenesis after hMSC culture. However, melanin synthesis was promoted after ascorbic acid treatment, which can be used to develop artifical skin for replacing real himan skin graft after further investigation.

( 출처 : SUMMARY 27p )

목차 Contents

• 표지 ... 1
• 제출문 ... 3
• 보고요약서 ... 6
• 요약문 ... 7
• SUMMARY ... 21
• CONTENTS ... 36
• 목차 ... 37
• 제 1 장 연구개발과제의 개요 ... 38
• 제 1 절 연구개발의 필요성 ... 38
• 제 2 절 연구개발의 목적 및 범위 ... 42
• 1. 연구개발 최종목표 ... 42
• 2. 연구개발의 범위 ... 42
• 제 2 장 국내외 기술개발 현황 ... 44
• 제 1 절 선진국 연구개발 동향 및 기술수준 ... 44
• 제 2 절 국내 연구개발 동향 및 선진국과의 기술격차 ... 47
• 제 3 장 연구개발수행 내용 및 결과 ... 50
• 제 1 절 방사선 이용 구강점막 부착용 하이드로겔 제조기술 개발 ... 50
• 제 2 절 방사선 이용한 약물 함유 점막 부착용 하이드로겔개발 ... 56
• 제 3 절 방사선을 이용한 세포배양용 지지체 제조 기술 개발 ... 62
• 제 4 절 인공피부 모델 형성 기술 개발 ... 66
• 제 5 절 인공피부 모델의 특성 평가 ... 71
• 제 4 장 목표달성도 및 관련분야에서의 기여도 ... 74
• 제 1 절 목표달성도 ... 74
• 1. 1 차년도 연구목표 ... 74
• 2. 2차년도 연구목표 ... 75
• 3. 3차년도 연구목표 ... 78
• 제 2 절 관련분야에서의 기여도 ... 81
• 제 5 장 연구개발결과의 활용계획 ... 82
• 제 6 장 연구개발과제에서 수집한 해외과학기술정보 ... 84
• 제 7 장 참고문헌 ... 85
• 방사선 이용 구강점막 부작용 약물 전달체 제조 기술 개발 ... 88
• 제출문 ... 89
• 보고요약서 ... 91
• 요약문 ... 92
• SUMMARY ... 100
• CONTENTS ... 107
• 목차 ... 108
• 제 1 장 연구개발과제의 개요 ... 109
• 제 1 절 연구개발의 필요성 ... 109
• 제 2 절 연구개발의 목적 및 범위 ... 111
• 1. 연구개발 최종목표 ... 111
• 2. 연구개발의 범위 ... 111
• 제 2장 국내외 기술개발 현황 ... 112
• 제 1 절 선진국 연구개발 동향 및 기술수준 ... 112
• 제 2절 국내 연구개발 동향 및 선진국과의 기술격차 ... 114
• 제 3장 연구개발수행 내용 및 결과 ... 115
• 제 1 절 방사선 이용 구강 점막 부착용 하이드로겔 제조 기술 개발 ... 115
• 1. PAA 이중구조의 구강점막 제형 제조 기술 개발 ... 115
• 2. 점막 접착력이 향상된 PAA/PEG 하이드로겔 개발 ... 122
• 3. 구강점막 접착력 향상을 위한 PVA/PAA 하이드로겔 개발 ... 132
• 4. PVA/PEG 하이드로겔의 열처리에 따른 영향성 평가 ... 137
• 5. PAA 나노겔 제조 기술 개발 ... 143
• 6. Ethyl cellulose 마이크로겔 개발 ... 149
• 제 2절 방사선 이용한 약물 함유 점막 부착용 하이드로겔개발 ... 152
• 1. 약물이 함유된 PAA의 효능 및 안정성 검증 ... 152
• 2. 은나노입자 함유 PVA 하이드로겔 개발 ... 162
• 3. 은나노입자 함유 PVA/PEG 하이드로겔 개발 ... 169
• 4. 은나노업자 함유 PVA/PEG 하이드로겔의 반응시간에 따른 특성 ... 174
• 5. CMC 하이드로겔에서 은나노입자 제조 ... 177
• 6. 산화아연 및 염화아연이 첨가된 PAA 하이드로겔 개발 ... 184
• 7. 구강점막 부착성 고분자를 이용한 약물의 가용화 및 서방화 ... 194
• 8. 구강점막 부착성 폴록사머 하이드로겔 제조 및 약물전달특성 분석 ... 200
• 제 4장 목표달성도 및 관련분야에서의 기여도 ... 217
• 제 1 절 목표달성도 ... 217
• 1. 1차년도 연구목표 ... 217
• 2.2차년도 연구목표 ... 217
• 3 . 3차년도 연구목표 ... 219
• 제 2 절 관련분야에서의 기여도 ... 221
• 제 5장 연구개발결과의 활용계획 ... 222
• 제 6장 연구개발과제에서 수집한 해외과학기술정보 ... 223
• 제 7 장 참고문헌 ... 224
• 방사선 이용 다기능 동물실험 대체용 인공피부 제조 기술 개발 ... 226
• 제출문 ... 227
• 보고요약서 ... 229
• 요약문 ... 230
• SUMMARY ... 237
• CONTENTS ... 246
• 목차 ... 248
• 제 1 장 연구개발과제의 개요 ... 249
• 제 1 절 연구개발의 필요성 ... 249
• 제 2절 연구개발의 목적 및 범위 ... 253
• 1. 연구개발 최종목표 ... 253
• 2. 연구개발의 범위 ... 253
• 제 2장 국내외 기술개발 현황 ... 256
• 제 1 절 선진국 연구개발 동향 및 기술수준 ... 256
• 제 2절 국내 연구개발 동향 및 선진국과의 기술격차 ... 258
• 제 3장 연구개발수행 내용 및 결과 ... 261
• 제 4장 목표달성도 및 관련분야에서의 기여도 ... 361
• 제 1 절 목표달성도 ... 361
• 1. 1차년도 연구목표 ... 361
• 2.2차년도 연구목표 ... 362
• 3.3차년도 연구목표 ... 364
• 제 2절 관련분야에서의 기여도 ... 366
• 제 5장 연구개발결과의 활용계획 ... 368
• 제 6장 연구개발과제에서 수접한 해외과학기술정보 ... 370
• 제 7 장 참고문헌 ... 371
• 끝페이지 ... 373