본 연구에서는 flavonoid apigenin과 baicalein의 세포 실험으로 콜라겐 회복효과를 확인하고 전사체 비교 분석을 통하여 collagen 회복 효과를 뒷받침할 수 있는 결과를 제안하고자 한다. 연구 결과 apigenin과 baicalein은 HS68 세포주에서 UV에 의해 손상된 I형 collagen 회복에 효과를 보였으며, 두 물질 모두에서 ECM 관련 유전자들의 발현 변화를 확인하였다. ECM을 조절하는 공통적인 메커니즘을 보일 뿐 아니라 각각 다른 범주의 유전자들을 변화시키는 결과도 보여서, 세포에 다양한 영향을 미치는 것으로 예측할 수 있었다.

혈관내큰B세포림프종은 악성 림프구의 혈관 내 성장을 보이는 드문 질환으로, 말초혈액 또는 혈관 외 종괴를 보이지 않는다. 이 림프종은 빠른 파종 및 공격적 성향 때문에 나쁜 예후를 가진다. 그러나 질병특유소견이 없어 진단이 어려운 실정이다. 저자들은 십이지장 위장관기질종양의 수술 검체 내에서 혈관내큰B세포림프종이 진단된 증례를 발견하였기에 문헌고찰과 함께 보고하는 바이다.

최근 조직공학 기술의 발달로 재해와 질병으로 인한 손상된 조직과 장기의 대체연구들 이 진행되고 있다. 장기 대체 연구의 핵심 요소 중 하나는 재건된 조직이나 장기가 혈관 망을 형성하여 host tissue로부터 양분과 산소의 전달이다. 본 연구는 조직공학 기법을 이용하여 장기 재건에 필수적인 혈관 재건을 위해 혈관을 구성하는 주세포인 endothelial cell을 체외에서 배양하는 것이다. Endothelial cell(EC)배양을 위해서는 세포지지체인 세 포외 기질(External cellular metrics, ECM)을 필요로 하기 때문에 ECM중에 대표적인 collagen과 gelatin을 사용하여 지지체에 따른 체외배양능을 비교하였다. 실험 동물로는 돼지 대동맥을 채취하여, 대동맥 속에 collagenase type I을 주입하고, 혈관의 입·출구를 봉합한 상태로 10분 간 37℃에서 처리하였다. 관류된 용액은 10% FBS가 함유된 기본배 양액(EGM-2 media)을 사용하여 2번 수세한 후 회수된 세포를 각각의 ECM이 처리된 dish위에서 배양 하였다. EC세포인지를 확인하기 위해서 EC표지 인자인 CD31과 vWF 항체의 발현을 flow cytometry로 확인 하였고, 회수된 세포에서 두 단백질이 모두 발현 되었다. ECM에 따른 EC의 세포 형태를 비교하였을 때 형태학적 차이는 없었다. Basement Membrane Extract위에서 calcein-AM으로 염색된 EC는 ECM의 종류와 상관 없이 2-6시간 사이에 Tube Formation을 보였다. 또한 endothelial cell의 표지 마크인 CD31, Flk1, vWF의 mRNA 발현양과 IHC에 의한 단백질 발현을 조사한 결과 collagen 지지체 위에서 배양된 endothelial cells에서 발현양이 더 높았다. 결론적으로 두 가지 ECM에서 모두 성공적으로 endothelial cell의 배양이 가능하지만 collagen위에서 배양된 endothelial cell이 더 우수한 maintenance능력을 가짐을 확인 할 수 있었다.

Although the mechanism of the abnormal calcification in the calcifying odontogenic cyst (COC) was not elucidated so far, it has been known that the ghost cells are closely related to the calcification, producing dystrophic globular cementum-like materials, comparable to pilomatricoma in epithelium1). Here, we presented a case of COC occurred in left maxillary canine area of 23 years old female, exhibiting a collection of aberrant ossification admixed with basophilic ghost cells in comparison with seven cases of COC. In the polarizing microscope observation with Masson trichrome stain the present case clearly disclosed the typical birefringence of bony tissue, stained red in von Gieson stain, indicating the collagenous backbone. Some ghost cells showed the features of interdigitating epithelial attachments, empty spaces of nuclei, and reticular basophilic cytoplasms, which were similar to the basophilic ghost cells of philomatricoma. The present case demonstrated the aberrant ossification by basophilic ghost cells in COC similar to the ossification of pilomatricoma

The pattern of wound healing process differs markedly according to the cell types. Gingival wounds heal more rapidly without scar, however dermal wounds show collagen laid down in thick disorganized patterns and keloid formation. This h as b een s uggested t o be d ue t o the presence of d ifferent E C M components a nd c ytokines a s well a s growth factors. The purpose of this study was to examine the differential expression of genes in connection with keloid formation in gingival fibroblasts (hGFs) and dermal fibroblasts (hDFs) in response to inflammation. In this study, we investigated the differences between hGFs and hDFs in the expression and production of cyclooxygenase (COX-2), prostaglandins E2 (PGE2), transforming growth factor (TGF)-β, collagens, matrix metalloproteinases (MMPs), and tissue inhibitors of matrix metalloproteinases (TIMPs) which play important roles in collagen deposition in wound healing. The hGFs and hDFs were primary cultured and allocated to arachidonic acid (AA) treatment group and control group. Protein and mRNA were extracted right after (0 hr) and 24 hr after AA treatment. At a defined concentration of AA in hGFs and hDFs, MTT assay was performed. The mRNA and protein expression levels of COX-2, TGF-β, collagen 1 and 3, MMP 1 and TIMP 1 were examined by Real-time PCR and Western blots. The amounts of PGE2 were measured by enzyme-linked immunosorbent assay (ELISA).The expression of COX-2 and TGF-β exhibited reduced levels in hGFs , but were increased in hDFs at 24 hr after AA treatment. Production of PGE2 was increased in hGFs and hDFs at right after AA treatment but, not changed at 24 hr after AA treatment. The protein and mRNA expression of collagen 1 and 3 were decreased in hGFs , whereas increased in hDFs at 24 hr AA treatment. Expression of MMP-1 protein was increased in hGFs at 24 hr but, was decreased in hDFs at 24 hr compared with that of control. The protein expression of TIMP-1 was decreased in hGFs but, was increased in hDFs at 24 hr compared with that of control. These observations demonstrate differential expression between gingival and dermal fibroblasts in regulation of collagenolytic capacity by extracellular matrix-associated genes in keloid formation associated with wound repair.

Cementum is a hard connective tissue, produced by cementoblasts during tooth root formation, which provides for the attachment of the periodontal ligament to the roots and surrounding alveolar bone. Establishment of this attachment is an important event in the regeneration of lost periodontal tissues. We examined whether or not odontoblast conditioned media(CM) have a regulatory influence on the differentiation and mineralization of cementoblasts(murine cementoblastic cell line, OCCM-30) in vitro. To identify the effect of odontoblast conditioned media and dentin non collagenous proteins (dNCPs) on cementoblast differentiation and mineralization, we treated CM and dNCPs to cementoblast then differentiated the cells for 14 days. To evaluate the formation of mineralized nodules alizarin-red S staining was performed at 0,4,7 and 14 days. Expression of cementum matrix genes was measured by RT-PCR. Mineralization of cementoblasts was accelerated with CM from odontoblastic MDPC-23 and OD-11. The expression of BSP, ALP, and OC mRNA in cementoblastic OCCM-30 cells was facilitated by the MDPC-23 and OD-11 cells. The extracted dNCPs had little influence on the proliferation, cell cycle modification, and chemotaxis of OCCM-30 cells. Although the dNCPs did not exhibit chemotactic activities for cementoblasts, the dNCPs promoted the differentiation and mineralization of cementoblasts. In conclusion, the dentin matrix protein, or the secreted products of odontoblast, facilitates cementoblast differentiation and mineralization. This represents a new approach and suggests another avenue for cementum regeneration.

ISPARC (Secreted protein acidic and rich in cysteine) is detected in the bone stroma during wound-healing process. To understand the roles of SPARC in bony wound-healing process, SPARC cDNA were synthesized from rat calvarial osteoblast culture, and SPARC protein was synthesized from the cDNA. To observe the effects of SPARC protein on the differentiation of osteoblasts, bony defect were made on rat tibia, and the distributions of bone matrix related proteins and SPARC were investigated using immunohistochemistry. In the rat osteoblastic culture using untreated plastic surface, Collagen-SPARC treated surface presented higher protein synthesis than untreated surface or only collagen treated surface. SPARC synthesis in the bony defect of rat tibia was augmented by introducing SPARC to the bony defect. SPARC synthesis were increased from the center of the defect compared to the control. SPARC synthesis in cells of the center of the defect was increased and maintained for 14 days. We could conclude that SPARC introduction may affect the early bone matrix formation, including SPARC, and mineralization in bony wound healing process.

It has been reported that light-emitting diodes(LED) can be used in the treatment of oral diseases. Although bio-stimulatory effects of LED irradiation such as promotion of wound healing have been well known, there are few reports about molecular mechanisms associated with wound healing by LED irradiation. The purpose of the present study was to investigate the expression pattern of various extracellular matrix(ECM) molecules in relation to wound healing after LED irradiation on primary human gingival fibroblasts(hGFs) in vitro. The source of light for irradiation was a continuous-wave LED emitting at a wavelength of 635 nm, and manufactured that energy density was 5 mW/cm2 on sample surfaces. The hGFs were irradiated for 1 hour at 37℃ in 5% CO2 humidified chamber. Experimental samples were acquired at 0 (right after irradiation), 24 and 48 hour after irradiation. To investigate the molecular mechanisms associated with wound healing, we examined the mRNA expression of 6 types of collagens, 7 types of matrix metalloproteinases(MMPs) and 4 types of tissue inhibition of metalloproteinases(TIMPs) after LED irradiation by RT-PCR. The mRNA expression of collagen 4, MMP-3, 9, and 16, and TIMP-3 was influenced by LED irradiation. Generally, the collagen expression of the irradiation group was slightly increased, particularly collagen 4 was significantly increased at 0 hour. The expression of MMP-3 was increased at 0 and 24 hours and MMP-16 was increased at 24 hours, respectively. The expression of MMP-9 was decreased at 0 hour and increased at 24 and 48 hours. The mRNA expression of TIMP-3 was significantly decreased at 24 and 48 hours after irradiation. These results suggest that the altered expression of ECM molecules after LED irradiation may contribute to the accelerated wound healing.

본 연구는 SPARC(osteo n ect in) 이 석회화 물질의 형성에 밀접한 연관을 가진다는 짐 애 착안하여 골조직 재생 괴정 중의 기질-세포 상호자용에 있어 SPARC의 역할 석회화 조직 형성과 연관된 SPARC의 발현조절과 기능을 규명하고자 시행되었다 l꾀서 태자 두개관 배양세포로부터 SPARC cDNA를 합성하였으며 이를 이용하여 백서 SPARC 단백질을 합성하였다 SP때C 단맥 질을 이용해 백서 조골 세 포의 증식 에 미치는 영호노을 관찰하였다 동시 에 백서 장골에 인위적인 골결손을 형성하고 형성과정에서의 골기질 관련인자의 분포를 관찰하고 SPARC 의 분포를 in situ hyb ridiz a tion법을 이용하여 관찰하였다 백서에서 분리 합성된 cDNA 조각은 1231bp 크기 였으며 40kD 크기의 단백질을 발현하였다 백서 태자 두개관 세 포의 증식에서 SPARC과 콜라겐 으로 처리된 표띤은 콜리겐 단독으로 표면처 리된 t111 양표연보다 증가힌 세포증식과 단백질 합성 콜라겐 합성을 나티내 었다 백서 -?I-골이1 형성힌 골조직 지| 생괴정에서 SPARC을 투여힌 백서 ;<J-골의 경우 결손부 전체에 걸쳐 증가한 SPARC의 합성을 보였디 SPARC이 투여된 군에서는 질손부의 중심 에서부터 SPARC과 콜라겐의 합성이 증기한 양상을 나타내었으며 대조군과 뚜렷한 차이를 나타내었다 in siLu h yb ricli za Li o n 깨 을 이용한 관칠 에서 골결손부 중잉에 있는 세 포에서의 SPARC 발현이 증가하였으며 그 싱패는 1 4 일 까지 대 조군에 비해 높게 니티났디 이싱의 연구 에서 얻은 결론은 디음과 같다 SPARC은 콜라겐과 결합하여 백서태자 두개관세 포의 배양에서 세 포증식 및 단백질 한성 ‘ 콜라겐 합성 의 뚜렷한 증가를 나타내었으며 . 이 같은 OJ상은 SPARC 단독으로 표연 처리하였을 때 는 관찰되지 않았다. SPARC의 골절손부 투여는 골조직 의 재생을 촉진허며 초기 세 포외 기질의 합성을 증가시켰으며 특히 SP뻐C의 형성이 초기애 증가하는 앙싱 융 나타내었다 이상 의 결과로 볼 떼 SPARC의 투여는 골조직 손상회복 시 초기 골기질 형성과 석회화에 영향을 미치며 SPARC 자치1 의 형성 증가에도 영향 을 미치는것으로보인다

본 연구는 phosphatidylinositol 대사의 기질 및 억제물질이 돼지 난모세포의 체외 성숙·수정에 미치는 영향에 대하여 실험을 하였다. 난모세를 inositol (250 mM) 첨가 또는 무첨가한 mTLP-PVA 배양액에서 46시간 체외배양하였다. 성숙이 완료된 난모세포는 신선 돼지정액을 이용하여 6시간 동안 mTALP-PVA 배양액에서 체외수정을 시켰다. 수정 후 6시간에 이들 난모세포를 10% 혈청을 첨가한 TCM-199 배양액에서 추가로

To investigate the effect of extracellular matrix proteins on the in vitro development of ethanol-induced parthenogenetic eggs of ICR strain mice, those were cultured in vitro in fibronectin, gelatin, or collagen precoated culture dishes containing 1.5 ml of NaH-C03-BMOC-3 medium at 37 for 96 hrs. under the atmosphere of 5% and 95% air. Fibronectin, gelatin, or collagen significantly(P1.4, 45.4i1.4, and 44.8O.9, respectively. And the diameter of those eggs ranged 104.61.9, 102.82.3, and 103.4O.8 m, respectively.

조혈세포의 주요 서식지가 되는 골수기질세포는 줄기세포의 운영을 결정하는 다양한 인자들을 제공한다. 방사선 요법은 항암치료법으로 널리 활용되고 있으나, 조혈세포의 파괴로 인한 부작용이 심각한 문제로서 조혈세포에 의한 혈액 세포가 빠른 시간 내에 회복되는 것이 필수적이다. 본 연구에서는 방사선을 조사했을 때의 줄기세포 서식지를 구성하는 세포인 골수기질세포에서 발현되는 유전자를 탐색하여 그 기능과 조절 및 혈액 형성을 이해하는 기초를 마련하고자 하였다. 방법