• Development and Reproduction
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• v.17 no.4
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• pp.299-309
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• 2013

Transforming growth factor (TGF) family is well known to induce the chondrogenic differentiation of mesenchymal stem cells (MSC). However, the precise signal transduction pathways and underlying factors are not well known. Thus the present study aims to evaluate the possible role of C2 domain in the chondrogenic differentiation of human mesenchymal stem cells. To this end, 145 C2 domains in the adenovirus were individually transfected to hMSC, and morphological changes were examined. Among 145 C2 domains, C2 domain of protein kinase C eta ($PKC{\eta}$) was selected as a possible chondrogenic differentiation factor for hMSC. To confirm this possibility, we treated $TGF{\beta}3$, a well known chondrogenic differentiation factor of hMSC, and examined the increased-expression of glycosaminoglycan (GAG), collagen type II (COL II) as well as $PKC{\eta}$ using PT-PCR, immunocytochemistry and Western blot analysis. To further evaluation of C2 domain of $PKC{\eta}$, we examined morphological changes, expressions of GAG and COL II after transfection of $PKC{\eta}$-C2 domain in hMSC. Overexpression of $PKC{\eta}$-C2 domain induced morphological change and increased GAG and COL II expressions. The present results demonstrate that $PKC{\eta}$ involves in the TGF-${\beta}3$-induced chondrogenic differentiation of hMSC, and C2 domain of $PKC{\eta}$ has important role in this process.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.11 no.9
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• pp.3358-3365
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• 2010

This study was carried out to investigate the effect of Sorbus commixta (SC), Geranium thunbergii (GT) and their mixture (SC:GT=1:1, MIX) on inhibition of bone loss and chondral defect. To examine their activities, we measured the alkaline phosphatase (ALP) activity in human osteoblast-like MG-63 cells and performed tartrate-resistant acid phosphate (TRAP) staining in osteoclast differentiated from Raw264.7 cells. To investigate the influence on chondrocyte differentiation, we performed alcian-blue staining in chondrocyte differentiated from ATDC5 cells. All of SC, GT and MIX did not increase ALP activity in MG-63 cells. However, SC and mixture (SC:GT=1:1, MIX) significantly inhibited osteoclastic differentiation. And they also induced chondrocyte differentiation. These results suggest that SC and GT may have a potential for the treatment of bone loss and chondral defect by suppression of osteoclast differentiation and stimulation of chondrocyte differentiation. Therefore, clarification of their mechanisms and active components will be needed.

• Journal of Korean Foot and Ankle Society
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• v.25 no.4
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• pp.149-156
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• 2021

Purpose: Given the lack of definite evidence-based guidelines in clinical practice, there may be a wide variation in treatment protocols for osteochondral lesions of the talus (OLT). Based on the Korean Foot and Ankle Society (KFAS) member survey, this study aimed to report the current trends in the management of OLT. Materials and Methods: A web-based questionnaire containing 30 questions was sent to all KFAS members in September 2021. The questions were mainly related to clinical experience and preferences in diagnosis, conservative, and surgical treatments for patients with OLT. Answers with a prevalence of ≥50% of respondents were considered a tendency. Results: Sixty-two (11.3%) of the 550 surgeons queried responded to the survey. The responses to 9 (30.0%) of the total of 30 questions established a tendency. Answers exhibiting a tendency were as follows; additional diagnostic tools except for plain radiograph (magnetic resonance imaging), most common conservative treatment method (oral medication, rest), most important radiological factor in decision making for surgical treatment and method (size of the lesion, ankle instability, loose bodies), most important patient factors in decision making for surgical treatment and method (age, activity or occupation), infrequently requiring posterior arthroscopy (less than 3%), most common revision surgery for failed bone marrow stimulation procedure (osteochondral autograft transplantation [OAT]), not requiring additional procedure for donor site in OAT, the main reason for unsatisfactory result after OAT (persistent pain without radiological abnormality), no generalization of autologous chondrocyte implantation or chondrogenesis using stem cells. Conclusion: This study presents updated information on current trends in the management of OLT in Korea. Both consensus and variations in the approach to patients with OLT were revealed through this survey. Since recent biologic efforts to regenerate cartilage have been unsuccessful, further studies to identify clinical evidence would be needed.

• Journal of the korean academy of Pediatric Dentistry
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• v.30 no.4
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• pp.643-653
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• 2003

Fibroblast growth factor (FGF) / FGF receptor (FGFR) mediated signaling is required for skeletogenesis in cluding intramembranous and endochondral ossifications Runx2 ($Cbfa1/Pebp2{\alpha}A/AML3$) is an essential transcription factor for osteoblast differentiation and bone formation. Murine calvaria and mandible are concurrently undergoing both intramembranous bone and cartilage formations in the early developmental stage. However the mechanism by which these cartilage formations are regulated remains unclear. To elucidate the effect of FGF signaling on development of cranial sutural cartilage and Meckel's cartilage and to understand the role of Runx2 in these process, we have done both in vivo and in vitro experiments. Alcian blue staining showed that cartilage formation in sagittal suture begins from embryonic stage 16 (E16), Meckel's cartilage formation in mandible from E12. We analyzed by in situ hybridization the characteristics of cartilage cells that type II collagen, not type X collagen, was expressed in sagittal sutural cartilage and Meckel's cartilage. In addition, Runx2 was not expressed in Meckel's cartilage as well as sagittal sutural cartilage, except specific expression pattern only surrounding both cartilages. FGF signaling pathway was further examined in vitro. Beads soaked in FGF2 placed on the sagittal suture and mandible inhibited both sutural and Meckel's cartilage formations. We next examined whether Runx2 gene lies in FGF siganling pathway during regulation of cartilage formation. Beads soaked in FGF2 on sagittal suture induced Runx2 gene expression. These results suggest that FGF signaling inhibits formations of sagittal sutural and Meckel's cartilages, also propose that FGF siganling is involved in the proliferation and differentiation of chondroblasts through regulating the transcription factor Runx2.

• Journal of Veterinary Clinics
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• v.28 no.5
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• pp.486-496
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• 2011

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.