The purpose of this experiment was to examine the histological changes and the pattern of expression of type I, II collagen in the elongated area by distraction osteogenesis in the rabbit mandible. Sixteen rabbits weighing 2.5kg-3kg were used for this experiment. Experimental group was distracted at the rate of 0.7mm, twice/day for 7days, and control group was only osteotomized. After 5 days latency, osteotomic site is distracted for 7days. Consolidation period is 28days. The animal was sacrificed at the 3rd, 7th, 14th, 28th day after the operation. The distracted bone was examined by histological analysis and RT-PCR analysis. The results were summarized as follows: 1. Experimental group was observed that the gaps between the distracted bone edges were occupied by new bone. 2. Expression of Type I collagen were detected throughout the experiment in both groups and Expression of Type I collagen were markedly increased during distraction and consolidation period in experimental group than control group. 3. Expression of Type II collagen were detected throughout the experiment in both groups and expression of Type II collagen were maintained at high level during distraction and consolidation period in experimental group than control group. From these results, in contrast to type II collagen, type I collagen seemed to be more expressed by mechanical stimuli during distraction and consolidation period. The predominent mechanism of new bone formation in the distraction gap was intramembranous bone formation, but some of the regenerated bone was formed by endochondral ossification.
Animals ; Collagen ; Collagen Type I ; Collagen Type II* ; Mandible* ; Osteogenesis ; Osteogenesis, Distraction* ; Rabbits

No Abstract Available.
Cell Adhesion Molecules* ; Cell Adhesion* ; Collagen Type II*

PURPOSE: We intended to check the growth rates and phenotypic markers of chondrocytes in the dedifferentiated cells cultivated in various conditions in order to establish the ideal culture system for implantation. MATERIALS AND METHODS: Culturing rabbit chondrocytes from proximal tibia, we checked the phenotypes at first, second, and third week. Then we cultured the chondrocytes in different circumstances such as monolayer or three dimensional gel in the presence or abscence of TGF-B1, and checked the growth rates and phenotypic markers. RESULTS: There was no difference in growth rates and mRNA level of type I, type II collagen and aggrecan between the cells cultured in monolayer and three dimensional gel of collagen. However, the responses of the cells to TGF-B1, were quite different between these two groups. In monolayer culture, the expression of type I collagen was depressed by TGF-B1 while the growth rate was markedly increased. Oppositely in three dimensional culture, the mRNA level of type I collagen was markedly increased and the growth rate was completely suppressed by TGF-B1. The expression of type II collagen could be detected only in TGF-B1-treated cells cultured in three dimensional gel for 4 or more days. The mRNA level of aggrecan was also increased by TGF-B1, in the cells cultured in three dimensional gel. CONCLUSIONS: These results suggest that the number of chondrocytes can be efficiently expanded by culturing the cells in monolayer and the phenotypes of chondrocyte can be restored by culturing the cells in three dimensional gel containing TGF-B1. The application of semi-solid gel containing differentiated chondrocytes in physeal implantation should be further evaluated
Aggrecans ; Chondrocytes* ; Collagen ; Collagen Type I ; Collagen Type II ; Phenotype ; RNA, Messenger ; Tibia ; Transforming Growth Factor beta1*

BACKGROUND: Cartilage tissue engineering (CTE) aims to obtain a structure mimicking native cartilage tissue through the combination of relevant cells, three-dimensional scaffolds, and extraneous signals. Implantation of ‘matured’ constructs is thus expected to provide solution for treating large injury of articular cartilage. Type I collagen is widely used as scaffolds for CTE products undergoing clinical trial, owing to its ubiquitous biocompatibility and vast clinical approval. However, the long-term performance of pure type I collagen scaffolds would suffer from its limited chondrogenic capacity and inferior mechanical properties. This paper aims to provide insights necessary for advancing type I collagen scaffolds in the CTE applications. METHODS: Initially, the interactions of type I/II collagen with CTE-relevant cells [i.e., articular chondrocytes (ACs) and mesenchymal stem cells (MSCs)] are discussed. Next, the physical features and chemical composition of the scaffolds crucial to support chondrogenic activities of AC and MSC are highlighted. Attempts to optimize the collagen scaffolds by blending with natural/synthetic polymers are described. Hybrid strategy in which collagen and structural polymers are combined in non-blending manner is detailed. RESULTS: Type I collagen is sufficient to support cellular activities of ACs and MSCs; however it shows limited chondrogenic performance than type II collagen. Nonetheless, type I collagen is the clinically feasible option since type II collagen shows arthritogenic potency. Physical features of scaffolds such as internal structure, pore size, stiffness, etc. are shown to be crucial in influencing the differentiation fate and secreting extracellular matrixes from ACs and MSCs. Collagen can be blended with native or synthetic polymer to improve the mechanical and bioactivities of final composites. However, the versatility of blending strategy is limited due to denaturation of type I collagen at harsh processing condition. Hybrid strategy is successful in maximizing bioactivity of collagen scaffolds and mechanical robustness of structural polymer. CONCLUSION: Considering the previous improvements of physical and compositional properties of collagen scaffolds and recent manufacturing developments of structural polymer, it is concluded that hybrid strategy is a promising approach to advance further collagen-based scaffolds in CTE.
Cartilage* ; Cartilage, Articular ; Chondrocytes ; Collagen Type I ; Collagen Type II ; Collagen* ; Extracellular Matrix ; Mesenchymal Stromal Cells ; Polymers ; Tissue Engineering*

BACKGROUND: Changes in the composition of the extracellular matrix (ECM) occur between the proliferating and involuted phases of infantile hemangiomas (IH), and are associated with angiogenic growth. We examined the composition of the ECM in proliferating and involuted IHs and assessed correlations between the composition of the ECM and whether the IH was in the proliferating or the involuted phase. METHODS: We evaluated IH samples from a cohort of patients who had five proliferating IHs and five involuted IHs. The following ECM molecules were analyzed using enzyme-linked immunosorbent assays and immunohistochemistry: laminin, fibronectin, collagen type I, collagen type II, and collagen type III. RESULTS: The involuted IHs had higher levels of deposition of collagen type III than the proliferating IHs. The median values (interquartile ranges) were 1.135 (0.946-1.486) and 1.008 (0.780-1.166) (P=0.019), respectively. The level of laminin was higher in involuted IHs than in proliferating IHs, with median values (interquartile ranges) of 3.191 (2.945-3.191) and 2.479 (1.699-3.284) (P=0.047), respectively. Abundant collagen type III staining was found in involuted IHs. Laminin alpha4 chain staining was clearly present within the basement membrane adjacent to the blood vessels, and was significantly more intense in involuted IHs than in proliferative IHs. CONCLUSIONS: Involuted hemangiomas showed extensive deposition of collagen III and laminin, suggesting that differences in the composition of the ECM reflect stages of the development of IHs. This pattern may be due to the rapid senescence of IHs.
Aging ; Basement Membrane ; Blood Vessels ; Cohort Studies ; Collagen ; Collagen Type I ; Collagen Type II ; Collagen Type III ; Enzyme-Linked Immunosorbent Assay ; Extracellular Matrix* ; Fibronectins ; Hemangioma* ; Humans ; Immunohistochemistry ; Laminin

PURPOSE: To evaluate the usefulness of three dimensional culture using alginate-fibrin beads on the chondrogenic differentiation of rabbit's perichondrial cells. MATERIALS AND METHODS: Rib perichondrial cells from rabbit expanded by monolayer culture were cultured in monolayer, in alginate bead, and in alginate-fibrin beads. Reverse transcription polymerase chain reaction (RT-PCR) for type I, II, X collagen, and aggrecan was performed at 3 weeks after culture. At that time, we removed the alginate component from the alginate-fibrin bead. Then, the cell-fibrin beads were transplanted into the partial physeal defect of proximal tibia, and some beads were cultured on for additional 3 weeks for RT-PCR. Histologic examination was performed at 2, 4, 8, 12, and 16 weeks after operation. RESULTS: At 3 weeks, type II collagen gene expression was maintained regardless of the culture system used. However, at 6 weeks it was maintained only in three-dimensional culture system using alginate beads and fibrin beads. Histologic examination showed that the implanted perichondrial cell-fibrin beads formed small nest composed of chondrocyte-like cells and matrix. CONCLUSION: These results suggest that alginate-fibrin beads may be used as a biodegradable scaffold for cartilage engineering using perichondrial cells.
Aggrecans ; Cartilage ; Collagen ; Collagen Type II ; Fibrin ; Gene Expression ; Polymerase Chain Reaction ; Reverse Transcription ; Ribs ; Tibia

PURPOSE: We had examined the effect of high dose MTX loaded acrylic cement in the treatment of type II collagen induced arthritis. MATERIALS AND METHODS: we used inbreed rat(16weeks - 20weeks) and divided into control group and experimental groups. inbreed rat was induced arthritis by prepared collagen through injection at the tail base. The control group was injected with collagen only. but the experimental groups were inserted intraperitoneally MTX-loaded acryl cement: 25mg, 20mg, 15mg, and 10mg MTX in acrylic cement (0.5gm) respectively. The histopathologic results were investigated to determine the effect of MTX on the 3rd, 7th, 14th, 28th, 42th, and 56th day after inserting high dose MTX loaded acrylic cement which was inserted intraperitoneally 2weeks later of collagenase injection RESULTS: The arthritis index of the MTX-loaded groups had reduced than arthritis index of the control group according to the dosage and In histopathological test, the experimental group injected with 15mg, 20mg, and 25mg of MTX had distinctive effects, compared to the control group, as dosage chronologically. CONCLUSION: The treatment of high-dose MTX loaded acrylic cement might be effective on rheumatoid arthritis managment.
Animals ; Arthritis* ; Arthritis, Rheumatoid ; Collagen ; Collagen Type II* ; Collagenases ; Methotrexate* ; Rats* ; Tail

PURPOSE: The purpose of this study is to identify nature of regenerated articular cartilage after microfracture surgery. MATERIALS AND METHODS: From Oct. 1997 to Nov. 1998, 40 knees were treated for osteoarthritis by arthroscopic microfracture technique. In the 18 knees, during the second arthroscopic procedure, biopsy specimens extending to the subchondral bone were taken and immunohistochemical staining was done to identify type of collagen. One patient was man and 17 patients were women. Average age of the patients were 58 years (range, 40-75 years). RESULTS: Type II collagen in articular cartilage appeared to be brown color with this staining. Degree of staining were +3 in 4 knees(22%), +2 in 2 knees(11%), +1 in 7 knees(39%) and trace in 5 knees(28%). CONCLUSION: Microfracture surgery restores the function of the joint by forming predominantly hyaline-like cartilage containing type II collagen. As analyzing amounts of type II collagen with an immunohistochemical staining in regenerated cartilage, we can presume the prognosis of regenerated cartilage tissue after microfracture surgery.
Biopsy ; Cartilage* ; Cartilage, Articular ; Collagen ; Collagen Type II* ; Female ; Humans ; Joints ; Knee ; Osteoarthritis ; Prognosis

PURPOSE: Rheumatoid arthritis is a chronic multisystemic disease involving joints. It has been difficult to explain why the inflammatory responses of the rheumatoid arthritis (RA) are mostly limited to the joints. To explain this localizing phenomenon, we hypothetized that the cartilage, which exists in the joints, maintaines the activated status of synovial membrane and provides local specific environment for the maintenance of inflammatory response of arthritis. MATERIALS AND METHODS: Synovial fibroblast were proliferated with and without the addition of cartilage tissues, chondrocytes, type II collagens to the growth media respectively. MTS assay (CellTiter 96(R) AQ One Solution Cell Proliferation Assay) was done at 1st, 3rd, 4th, 5th, 6th, 7th, 9th and 11th days to evaluate the proliferation of the cells. The data were assessed with one-way ANOVA and unpaired t-test. Results : In a group where 0.1 g/ml of cartilage tissues were added to the growth media, the synovial fibroblasts from the normal people and rhematoid arthritic patients showed higher amount of proliferation from the 3rd day compared to the control group. When 0.02 g/ml of cartilage tissues were added, the synovial fibroblasts from the normal people showed higher amount of proliferation from the 4th day and those from RA patients from the 3rd day compared to the control group (p< 0.05). There was no significant statistical difference according to the concentration of the cartilage tissues.In a group where chondrocyte extract was added to the growth media, the synovial fibroblasts from the normal people showed higher amount of proliferation from the 4th day and those from RA patients from the 3rd day compared to the control group (p< 0.05). In a group where type II collagens were added to the growth media, the synovial fibroblasts from normal people and RA patients showed no significant statistical differences compared to the control group. CONCLUSION: The cartilage tissue is important in maintenance of synovial fibroblast proliferation. Of the cartilage tissue, chondrocyte, not extracellular type II collagen, played a major role. But further study is needed about what component of the chondrocyte extract maintaines the proliferation of synovial fibroblast.
Arthritis ; Arthritis, Rheumatoid ; Cartilage* ; Cell Proliferation ; Chondrocytes ; Collagen ; Collagen Type II ; Fibroblasts ; Humans ; Joints ; Synovial Membrane

OBJECTIVE: Osteoarthritis is a disorder or group of disorders affecting synovial joint, characterized at the tissue level by degenerative, regenerative and reparative structural changes in cartilage, synovium and bone. The histopathological understanding of osteoarthritis would help to clear the pathogenesis and evaluate the activity and progression of the disease. We studied the histochemical distribution of collagen in the search for and accurate morphometric m~rker for osteoarthritic progression. METHODS: The immunohistochemical analysis by monoclonal antibodies to type I, II and III collagen, the Masson's trichrome stain and the Picrosirius stain and polarization microscopy were performed in 10 osteoarthritis cartilages and 1 normal control cartilage. RESULTS: 1)By the immunohistochemical analysis, type II collagen was diffusely expressed in both normal and osteoarthritis cartilage but the expression of type I and III collagen was only shown around chondrocytes in the osteoarthritis cartilage. 2) There was no difference in Masson's trichrome stain of the normal and osteoarthritis cartilage. 3) By the Picrosirius stain and polarization microscopy, there was less diffuse birefringence of collagen and increased perichondronal birefringence in the osteoarthritis cartilage in comparison to normal cartilage. CONCLUSIONS: The Picrosirius-polarization method may be considered one of the useful morphometriic methods for the progression of osteoarthrits.
Antibodies, Monoclonal ; Birefringence ; Cartilage* ; Chondrocytes ; Collagen Type II ; Collagen* ; Humans* ; Joints ; Microscopy, Polarization ; Osteoarthritis ; Synovial Membrane