연골막과 연령이 가토에 이개연골 공여부 재새에 미치는 영향

백승우,박상현,김장호,방유현 大韓成形外科學會 1996 Archives of Plastic Surgery Vol.23 No.4

Cartilage graft has been used for the aesthetic and reconstructive surgery of nose, ear, and reginoal contour. Usual donor sites are costal, septal, or ear cartilages. Although there are many studies about the cartilage growth and the perichondrial effect in cartilage graft,few studies of the cartilage regrowth of donor site have been reproted. This experimental study includes the role of the perichondrium and the age in cartilage regrowth of donor site. Rabbits were devided in two groups, 10 young ravvits(3-4weeks) and 10 adult rabbits (10months). From each rabbit, the cartilage with perichondrium was removed from one ear, and the denuded cartilage was removed from the opposite ear. The regrowth of cartilage tissue at each cartilage harvest site was examined after 2 months. The results are as follows: 1. In young rabbits, the cartilages grew an average of 1.84mm(23.1%). In the perichondrial-remanined donor sites, an average of 2.04mm(25.6%) and an average of 1.63mm(20.6%) in the perichondrial-stripped donor sites. 2. In adult rabbits, the cartilages grew an average of 0.29mm(3.7%). In the perichondrial-remained donor sites, average of 0.36mm(4.5%) and an average of 0.22mm(2.8%) in the perichondrial-stripped donor sites. 3. In histologic studies, the active proliferating cells increased more in young rabbits than in adult rabbits, and increased more in the perichondrial-remained donor sites than in the perichondrial-stripped ones. In young rabbits, the cartilages regrew more in the perichondrial-remained donor sites than perichondrial-stripped ones in young rabbits, but in adults, the cartilages grew little compared to young rabbits without the relation to presence of perichondrium. Therefore, the results of this study suggest that the age is more important factor on the cartilage regrowth of the donor sites and the perichondrium has effect on the cartilage regrowth of only young rabbits.

OP-1 이 첨가된 Lyophilized Cartilage Allografts를 이용한 관절연골결손의 재생

신영진,김태준 大韓成形外科學會 2000 Archives of Plastic Surgery Vol.27 No.2

Joint cartilage defects do not heal and lead to post traumatic arthritis or degenerative joint diseases. The clinical problem is that so far, we have been unable to induce true cartilage healing and regeneration. Experimental attempts at repairing joint cartilage defects with various means have usually led to the formation of cartilage-like fibro-hyaline scars. We have recently shown that treatment of lyophilized allografts with recombinant osteoinductive protein-1 (OP-1 also known as BMP 7) induced the regeneration of articular cartilage over the head of lyophilized hemijoint allografts. A standardized 3 mm circular defect was created on the articular surface of the knee joint of 12 NZW rabbits. The defect was into carried as full thickness down to subchondral bone. In a blinded and randomized fashion, the rabbits were divided into four groups. In group 1(n=4), the ulcer was left untreated; in group Ⅱ(n=3), the ulcer was reconstructed with a 3 mm disk of lyophilized cartilage allograft; in group Ⅲ(n=2), the allograft used was pre-treated with low dose (20㎍) OP-1, and in group 1V(n=3), the allograft used was pre-treated with high dose(100㎍) OP-1. The range of motion was normal in all groups. The gross appearance of the untreated defect was still depressed and appeared like a big crater. The lyophilized allograft has a level surface which was not smooth. In high dose OP-1 group, the defect was healed much better than the lower dose treated OP-1 group. Under the histologic examination, untreated ulcer group never heals to the cartilage of around defect, a fissure always remains. Lyophilized allograft and low dose OP-1 treated lyophilized allograft groups seem to inhibit healing and there is fibrous scar in ulcer. In the high dose OP-1 treated lyophilized allograft group, new cartilage heals very well all around of the defect margin without fissure. The cartilage has a normal sequence of cellular maturation. In conclusion, in this study of full thickness joint cartilage defects, we confirmed finding in the literature that defect heals with cartilage like tissue which does not fuse with the surrounding cartilage. We found that lyophilize allograft prevents healing of defect, while lyophilized allograft treated with high dose (100㎍) OP-1 seems to induce regeneration of articular cartilage that heals very well to the surrounding defect without any crack or fissure.

Analysis of the Development of the Nasal Septum according to Age and Gender Using MRI

김인상,정영준,이민영,이기일,김혜영 대한이비인후과학회 2008 Clinical and Experimental Otorhinolaryngology Vol.1 No.1

Objectives. This study was designed to evaluate the normal development of the nasal septum in Koreans using sagittal MRI for the valuable clinical information on septal procedures. Methods. Two hundred eighty patients who had their whole nasal septum visualized in the midline sagittal view were selected among the 3,904 patients with brain MRI from January, 2004 to December, 2006 at Dankook University Hospital. The patients who had a history of nasal septal surgery or nasal trauma were excluded. Following parameters are calculated and analyzed: lengths of bony and cartilage dorsum and septal cartilage-nasal bone overlap, total septal area, septal cartilage area and, the proportion of the cartilage area to septal area and the maximal harvestable cartilage for grafting were calculated using the PACTM program. Results. All the parameters were increased until adolescence. Thereafter, bony dorsal length, cartilage dorsal length, total dorsal length, total septal area and maximal harvestable cartilage for grafting have not changed significantly with age, while SC-NB overlap length, septal cartilage area, and proportion of the cartilage area to the total septal area were significantly decreased with age. The SC-NB overlap length was positively correlated with the septal cartilage area and the proportion of the cartilage area to the total septal area. Conclusion. The small septal cartilage area and its proportion to the total septal area were significantly correlated with a short overlap length of the septal cartilage under the nasal bone. Septal procedures should be carefully performed in the elderly due to the risk of incurring saddle nose. Objectives. This study was designed to evaluate the normal development of the nasal septum in Koreans using sagittal MRI for the valuable clinical information on septal procedures. Methods. Two hundred eighty patients who had their whole nasal septum visualized in the midline sagittal view were selected among the 3,904 patients with brain MRI from January, 2004 to December, 2006 at Dankook University Hospital. The patients who had a history of nasal septal surgery or nasal trauma were excluded. Following parameters are calculated and analyzed: lengths of bony and cartilage dorsum and septal cartilage-nasal bone overlap, total septal area, septal cartilage area and, the proportion of the cartilage area to septal area and the maximal harvestable cartilage for grafting were calculated using the PACTM program. Results. All the parameters were increased until adolescence. Thereafter, bony dorsal length, cartilage dorsal length, total dorsal length, total septal area and maximal harvestable cartilage for grafting have not changed significantly with age, while SC-NB overlap length, septal cartilage area, and proportion of the cartilage area to the total septal area were significantly decreased with age. The SC-NB overlap length was positively correlated with the septal cartilage area and the proportion of the cartilage area to the total septal area. Conclusion. The small septal cartilage area and its proportion to the total septal area were significantly correlated with a short overlap length of the septal cartilage under the nasal bone. Septal procedures should be carefully performed in the elderly due to the risk of incurring saddle nose.

Tracheal Cartilage Regeneration by Progenitor Cells Derived from the Perichondrium

( Myung Hee Yoon ),( Ji Hae Kim ),( Chul Ho Oak ),( Tae Won Jang ),( Maan Hong Jung ),( Bong Kwon Chun ),( Sang Joon Lee ),( Jeong Hoon Heo ) 한국조직공학·재생의학회 2013 조직공학과 재생의학 Vol.10 No.5

The regenerative potential of adult tracheal cartilage is very limited which makes repair of this cartilage extremely challenging. However, the perichondrium has a large quantity of progenitor cells that differentiate directly into chondrocytes. Bone morphogenetic protein (BMP) stimulates chondrogenesis by inducing the differentiation of progenitor cells into chondrocytes which makes it a potential therapeutic agent for cartilage regeneration. In this present study, we investigated the mesenchymal stem cell characterization of progenitor cells from the perichondrial tissue of tracheal cartilage and the efficiency of the perichondrium for tracheal cartilage regeneration by evaluating the effect of BMP on cartilage regeneration in perichondrial progenitor cells. Progenitor cells were isolated from the fibrous perichodrial tissue of tracheal cartilage and cultured for phenotypic characterization of mesenchymal stem cells. Next, a 0.3-0.5 mm gap was made in the mid-ventral portion of the tracheal cartilages. In the control group (n=5), the resulting gap was left untreated (without perichondrial replacement). In the perichondrial group (n=15), the gap was replaced by fibrous perichondrial tissue. We euthanized all mice in each group (n=5) at 2, 4, and 20 weeks after the perichondrial replacement and examined the replacement site microscopically. The immunohistochemistry for BMP-2 was performed in progenitor cells from perichondrial tissue. The primary cultured cells from fibrous perichondrial tissue were positive for CD29, CD90, and CD105, but negative for CD45. The progenitor cells were capable of multipotent differentiation towards adipogenic and osteogenic lineages. The control group showed no regenerated cartilage 2 weeks after injury. The experimental group demonstrated proliferation of progenitor cells between both stumps at 2 weeks and regenerated hyaline cartilage was observed at 4 and 20 weeks. Regenerated cartilage was observed as separated islets between the ends of the host cartilage stumps. The gaps were nearly filled with regenerated cartilage at 20 weeks. BMP-2 was expressed in both fibrous and cabial perichondria chondrogenesis. The progenitor cells from perichondrial tissue had mesenchymal stem cell-like features and might play a crucial role in tracheal cartilage regeneration and repair. Expression of BMP-2 may induce formation of structural cartilage in a tracheal injury model.

Intra-Articular Atelocollagen Injection for the Treatment of Articular Cartilage Defects in Rabbit Model

Suh Dong Sam,Yoo Ji Chul,Woo Sang Hun,Kwak Andrew S. 한국조직공학과 재생의학회 2021 조직공학과 재생의학 Vol.18 No.4

Background: Atelocollagen is widely recognized as a biomaterial for regenerative medicine because of its good compatibility and low antigenicity. Injury of the outermost layer of articular cartilage, known as the lamina splendens, can lead to osteoarthritis (OA) and eventually full-thickness cartilage loss. The intra-articular injection of atelocollagen has been designed to restore the cartilage layer and cartilage defects in OA joints. In this study, we investigated the efficacy of atelocollagen as a cartilage supplement for joint defects. Methods: In this study, we evaluated the therapeutic effects of atelocollagen in animals with cartilage defects. Femoral groove defects were artificially created in 12 male New Zealand white rabbits, which were treated with intra-articular injection of either atelocollagen (experimental) or normal saline (control). The results were observed 3, 6, 9, and 12 weeks following macroscopic and histological evaluations. Results: At 3 weeks, cartilage tissue was restored in the experimental group, whereas the control group did not show signs of restoration. At 12 weeks, defects in both groups were filled with regenerated tissue, but the experimental group displayed a morphologically better appearance. Histologically, the regenerated tissue in the experimental group showed statistically significant improvement compared to the control group, with a structure similar to that of normal articular cartilage. Conclusion: The results showed that the intra-articular injection of atelocollagen enhanced cartilage regeneration following rabbit patellar groove defects. Therefore, intra-articular injection of atelocollagen can be used as an effective supplement for joint defects. Background: Atelocollagen is widely recognized as a biomaterial for regenerative medicine because of its good compatibility and low antigenicity. Injury of the outermost layer of articular cartilage, known as the lamina splendens, can lead to osteoarthritis (OA) and eventually full-thickness cartilage loss. The intra-articular injection of atelocollagen has been designed to restore the cartilage layer and cartilage defects in OA joints. In this study, we investigated the efficacy of atelocollagen as a cartilage supplement for joint defects. Methods: In this study, we evaluated the therapeutic effects of atelocollagen in animals with cartilage defects. Femoral groove defects were artificially created in 12 male New Zealand white rabbits, which were treated with intra-articular injection of either atelocollagen (experimental) or normal saline (control). The results were observed 3, 6, 9, and 12 weeks following macroscopic and histological evaluations. Background: Atelocollagen is widely recognized as a biomaterial for regenerative medicine because of its good compatibility and low antigenicity. Injury of the outermost layer of articular cartilage, known as the lamina splendens, can lead to osteoarthritis (OA) and eventually full-thickness cartilage loss. The intra-articular injection of atelocollagen has been designed to restore the cartilage layer and cartilage defects in OA joints. In this study, we investigated the efficacy of atelocollagen as a cartilage supplement for joint defects. Methods: In this study, we evaluated the therapeutic effects of atelocollagen in animals with cartilage defects. Femoral groove defects were artificially created in 12 male New Zealand white rabbits, which were treated with intra-articular injection of either atelocollagen (experimental) or normal saline (control). The results were observed 3, 6, 9, and 12 weeks following macroscopic and histological evaluations. Results: At 3 weeks, cartilage tissue was restored in the experimental group, whereas the control group did not show signs of restoration. At 12 weeks, defects in both groups were filled with regenerated tissue, but the experimental group displayed a morphologically better appearance. Histologically, the regenerated tissue in the experimental group showed statistically significant improvement compared to the control group, with a structure similar to that of normal articular cartilage. Conclusion: The results showed that the intra-articular injection of atelocollagen enhanced cartilage regeneration following rabbit patellar groove defects. Therefore, intra-articular injection of atelocollagen can be used as an effective supplement for joint defects. Results: At 3 weeks, cartilage tissue was restored in the experimental group, whereas the control group did not show signs of restoration. At 12 weeks, defects in both groups were filled with regenerated tissue, but the experimental group displayed a morphologically better appearance. Histologically, the regenerated tissue in the experimental group showed statistically significant improvement compared to the control group, with a structure similar to that of normal articular cartilage. Conclusion: The results showed that the intra-articular injection of atelocollagen enhanced cartilage regeneration following rabbit patellar groove defects. Therefore, intra-articular injection of atelocollagen can be used as an effective supplement for joint defects.

가토 슬관절연골의 대기노출 시간에 따른 변성 및 재생

김익동,김풍택,박병철,최영욱,이상복 慶北大學校 醫科大學 1990 慶北醫大誌 Vol.31 No.2

Orthopedic procedures expose articular cartilage to operating room air for varying periods of time as long as two hours. Articular cartilage is dense white, glossy and smooth, and is composed of chondrocytes, collagen, and a hyaline matrix made up of proteoglycan molecules. Articular cartilage is a specialized tissue containing 75 to 85% water. Most of the water is trapped in a gel of matrix collagen and proteoglycan and is freely exchangeable with the synovial fluid and blood. Only about 6% of cartilage water is tightly bound. Theoretically, since cartilage gel is hyperhydrated, the water content is important in maintaining cartilage resiliency and lubrication of the articular surface. If a pathologic state is associated with reduction on water content, these properties may be altered, leading to impaired joint function. We reproduced the O.R. situation by exposing one knee of 24 mature New-Zealand white rabbits to ambient air (20-22℃) for 30 minutes to 2 hours, the opposite knee serving as a control. Following exposure, the joints were closed and the animal resumed cage activity for 1 day prior to sacrifice, at that time cartilage was removed from both exposed and control joints and prepared for gross, light microscopy and electron microscopy. We observed cartilages which were sacrificed at 1 week after operation, 2 weeks after and 4 weeks after for varying periods of exposing time in same method. The results were as follow; 1. A distinct color change was seen in the exposed cartilage in groups which were sacrificed postoperative at 1 day ; However minimal change was detected in each group sacrificed at 1 week & 2 weeks later, and no change was seen in each group sacrificed postoperative at 4 weeks. 2. Exposure to room air for 30 minutes produced chondrocyte necrosis in the superficial zone of rabbit articular cartilage. Exposure for 1 hour or longer produced chondrocyte necrosis up to radial zone. 3. Chondrocyte necrosis was not seen in each group for varying periods of air exposure microscopically, which sacrificed postoperative at 1 week. 4. Proteoglycan content was minimal at 1 week postoperatively in each group ; However proteoglycan content increased progressively with time. 5. Change in cellularity were consistent with cartilage damage. Seemingly, exposing articular cartilage to air for 2 hours caused reversible cartilage damage.

가토의 관절 연골 결손 부위에서 미세 골절과 골형성 단백-2가 연골의 치유에 미치는 영향

이승준,김학준,양희석,김병수,박정호 한국조직공학과 재생의학회 2009 조직공학과 재생의학 Vol.6 No.13

Articular cartilage has a limited capacity for repair. The lack of repair in partial cartilage injury may result from the avascular nature of cartilage. Several methods have been developed to accelerate cartilage healing. Microfracture is an operative method that can facilitate access to stem cells in bone marrow and stimulate cartilage regeneration. However, microfracture has been shown to result in regeneration with fibrocartilage. The bone morphogenetic protein-2 has been shown to induce the expression of cartilage and bone marker. These findings prompted us to investigate rhBMP-2’s ability to modulate the repair of full-thickness defect of articular cartilage. The full thickness articular cartilage defects of 6x3mm size were created in the trochlear groove of forty-eight rabbits. Twelve defects were left empty, and other thirty-six defects were treated with microfracture. Among those thirty-six defects, twelve defects were filled with fibrin glue, and twelve defects with fibrin glue and rhBMP-2. The animals were killed at 8 weeks after surgery. The repair tissue was examined histologically and evaluated with real time PCR to analysis collagen type. After microfracture at the cartilage defect, treatment with fibrin glue and rhBMP-2 accelerated cartilage healing and the formation of new subchondral bone, and showed more hyaline-like cartilage components than other groups on the histological findings. Real-time PCR showed higher amounts of collage type 1 and type 2 in the group filled with fibrin glue and rhBMP-2. Articular cartilage has a limited capacity for repair. The lack of repair in partial cartilage injury may result from the avascular nature of cartilage. Several methods have been developed to accelerate cartilage healing. Microfracture is an operative method that can facilitate access to stem cells in bone marrow and stimulate cartilage regeneration. However, microfracture has been shown to result in regeneration with fibrocartilage. The bone morphogenetic protein-2 has been shown to induce the expression of cartilage and bone marker. These findings prompted us to investigate rhBMP-2’s ability to modulate the repair of full-thickness defect of articular cartilage. The full thickness articular cartilage defects of 6x3mm size were created in the trochlear groove of forty-eight rabbits. Twelve defects were left empty, and other thirty-six defects were treated with microfracture. Among those thirty-six defects, twelve defects were filled with fibrin glue, and twelve defects with fibrin glue and rhBMP-2. The animals were killed at 8 weeks after surgery. The repair tissue was examined histologically and evaluated with real time PCR to analysis collagen type. After microfracture at the cartilage defect, treatment with fibrin glue and rhBMP-2 accelerated cartilage healing and the formation of new subchondral bone, and showed more hyaline-like cartilage components than other groups on the histological findings. Real-time PCR showed higher amounts of collage type 1 and type 2 in the group filled with fibrin glue and rhBMP-2.

The Effect of Cultured Perichondrial Cell Sheet Covered Highly Active Engineered Cartilage: in vivo Comparative Assessment

박세일,문영미,정재호,장광호,안면환 한국임상수의학회 2011 한국임상수의학회지 Vol.28 No.5

A special mesenchymal tissue layer called perichondrium has a chondrogenic capacity and is a candidate tissue for engineering of cartilage. To overcome limited potential for chondrocyte proliferation and re-absorption, we studied a method of cartilage tissue engineering comprising chondrocyte-hydrogel pluronic complex (CPC) and cultured perichondrial cell sheet (cPCs) which entirely cover CPC. For effective cartilage regeneration, cell-sheet engineering technique of high-density culture was used for fabrication of cPCs. Hydrogel pluronic as a biomimetic cell carrier used for stable and maintains the chondrocytes. The human cPCs was cultured as a single layer and entirely covered CPC. The tissue engineered constructs were implanted into the dorsal subcutaneous tissue pocket on nude mice (n = 6). CPC without cPCs were used as a controls (N = 6). Engineered cartilage specimens were harvested at 12 weeks after implantation and evaluated with gross morphology and histological examination. Biological analysis was also performed for glycosaminoglycan (GAG) and type II collagen. Indeed, we performed additional in vivo studies of cartilage regeneration using canine large fullthickness chondrial defect model. The dogs were allocated to the experimental groups as treated chondrocyte sheets with perichondrial cell sheet group (n = 4), and chondrocyte sheets only group (n = 4). The histological and biochemical studies performed 12 weeks later as same manners as nude mouse but additional immunofluorescence study. Grossly, the size of cartilage specimen of cPCs covered group was larger than that of the control. On histological examination, the specimen of cPCs covered group showed typical characteristics of cartilage tissue. The contents of GAG and type II collagen were higher in cPCs covered group than that of the control. These studies demonstrated the potential of such CPC/cPCs constructs to support chondrogenesis in vivo. In conclusion, the method of cartilage tissue engineering using cPCs supposed to be an effective method with higher cartilage tissue gain. We suggest a new method of cartilage tissue engineering using cultured perichondrial cell sheet as a promising strategy for cartilage tissue reconstruction. 조직공학적 인공연골재생에 대한 관심이 증가함에 따라 많은 연구들이 활발히 수행되고 있으나 임상적인 적용의 한계를 극복하기위한 고효능을 보유한 양질의 연골조직생산의 필요성이 증가되고 있다. 인공연골은 자연연골과는 달리 ‘연골막(perichondrium)’을 포함하고 있지 않기 때문에 장기간 생체 내에 삽입된 후에 서서히 흡수 또는 변형으로 임상적 활용에 한계가 있다고 있다. 이에 본 연구는 양질의 연골조직생산을 목적으로, 세포판 제작기법(cell sheet engineering technique) 을 기반으로 한 인체유래의 배양 연골막(cultured perichondrium)을 이용하여 만든 인공연골막세포판(cultured perichondrial cell sheet)의 생체 내 특성을 비교 분석하고, 배양된 연골막을 피복하여 고효능화를 유도한 인공연골복합체의 생체내 재생효능 및 조직특성을 비교 평가하고자 하였다. 본 연구에서는 thymic nude mouse 의 피하이식모델(study 1, n = 12)을 이용하여 담체로 hydrogel을 이용한 배양연골막 복합체의 생체내 효능을 분석하였고, 중대형동물의 대량연골 결손시의 재생효능을 평가하기 위하여 개의 무릎연골에 1 × 2 cm의 대량연골 결손모델(study 2, n = 12)을 통하여 인공배양세포판을 이식하였다. 이식12주 후 이식편을 회수하여 생화학, 분자생물학 및 면역조직학 분석을 시행한 결과, 배양연골막 복합체의 생체내 효능이 단독이식군에 비해 변형이나 과증식 없이 우수한 결과를 나타내었다. 본 연구의 결과로 토대로 배양연골막을 피복한 인공연골막의 관절내 효과를 규명하여 실제 임상적용을 조기화하는 기반을 제공하고 인공연골의 문제점이었던 변형과 흡수를 줄인 고효능 인공연골 제작기법을 제공하는데 유용할 것으로 기대된다.

인간의 초자 연골 세포와 탄성 연골 세포를 이용한 조직공학적 연골 형성

신동필,한은희,박재우,김신윤,김도원,서인수,임정옥,백운이,조병채 경북대학교 병원 2003 경북대학교병원의학연구소논문집 Vol.7 No.1

The purpose of this study was to evaluate the influence of different type of PLGA scaffolds on the formation of human auricular and septal cartilages. The scaffolds were formed in tubular shape from 110,000g/mol PLGA(poly lactic glycolic acid) and 220,000g/mol one Elastic cartilage was taken from the ear of a patient aged under 20 years old and hyaline cartilage from the nasal septum The chondrocytes cells were then isolated by Klausburn method After second passages, the chondrocytes were seeded on the PLGA scaffolds followed by in vitro culture for one week The cells-PLGA scaffold complex was implanted at the back of nude mouses for 8 weeks. The tissue engineered cartilages were separated from nude mouse and examined histologically after staining with the Hematoxylin Eosin and Verhoeff. The formation of extracellular matnx and the porosity of the scaffolds were examined by scanning electron microscopy The pores were well formed and uniformly distributed in both 110,000g/mol and 220,000g/mol one And hyaline cartilage was proliferated better in vitro culture than elastic cartilage After 8 weeks in vivo culture, cartilage was well formed with 110,000g/mol PLGA, however lumen was collapsed in contrast with 220,000g/mol PLGA scaffold, neocartilage was formed in minimal amount while the architecture of scaffold was well preserved. Elastic cartiiage seems to be better than hyaline one in terms of neocartilage formation. Form the analysis after Verhoeff staining the cartilages, the neocartilage from elastic cartilage was proved to be elastic cartilage. In summary, there was no significant difference between elastic cartilage and hyaline cartilage in their morphologies, proliferation rates and the degree of cartilage formation since they were tissue engineered, however marked difference was found in neocartilage formation and preservation of scaffold architecture between 110,000g/mol PLGA scaffold and 220,000 one From the present findings, it is concluded that the influence of scaffold materials is significantly higher than that of different types of cells on the formation of new tissues.

The Re-Expression of Collagen Type 2, Aggrecan and Sox 9 in Tissue-Engineered Human Articular Cartilage

( Munirah Sha`ban ),( Aminuddin Bin Saim ),( Samsudin Osman Cassim ),( Chua Kien Hui ),( Fuzina Nor Hussein ),( Ruszymah Bt Hj Idrus ) 한국조직공학과 재생의학회 2005 조직공학과 재생의학 Vol.2 No.4

This study was designed to verify the optimal · basic culture media that promote chondrocytes proliferation in vitro in order to facilitate adequate amount of chondrocytes for cartilage reconstructionas well as maintaining cartilage specific phenotype. Human articular chondrocytes were cultured in three types of basic culture media Ham`s F12, DMEM and the equivalent mixture of F12:DMEM. Cultured chondrocytes were trypsinized as they reached confluency. The viability and total number of cell were recorded at every passage. Large-scale culture expansion was used to reconstruct tissue-engineered cartilage. Quantitative RT-PCR analysis was used to evaluate the expression of collagen Type II, collagen Type I, aggrecan and Sox 9 gene, both in monolayer culture and in the engineered cartilage. The mixture of F12:DMEM promotes significantly greater (p<0.05) chondrocytes proliferation at every passage compared to the individual medium. Monolayer cultured chondrocytes exhibited down-regulation expression pattern of collagen Type II gene, aggrecan and Sox 9, whilst the expression of collagen Type I is up-regulated. Tissue-engineered cartilage morphologically and histologically resembled normal hyaline cartilage. Moreover, tissue-engineered cartilage re-expressed the specific chondrogenesis markers; collagen Type II, aggrecan and Sox 9. In conclusion, the mixture of F12:DMEM enhanced human articular chondrocytes proliferation thus provided adequate amount of chondrocytes for cartilage reconstruction. The new cartilage formed phenotypically resembles native cartilage. This results hold promise for the use of tissue-engineered cartilage implant for future orthopaedic reconstructive surgery.