OBJECTIVES: To compare the effect of three different cryoprotectants on basic stem cell characteristics for the possibility of using well defined, dimethyl sulfoxide (DMSO) and serum free freezing solutions to cryopreserve human Wharton's jelly-derived mesenchymal stem cells (WJMSCs) following controlled rate freezing protocol. METHODS: The mesenchymal stem cells isolated from human Wharton's jelly were cryopreserved using 10% DMSO, 10% polyvinylpyrrolidone (PVP) and a cocktail solution comprising of 0.05 M glucose, 0.05 M sucrose and 1.5 M ethylene glycol following controlled rate freezing protocol. We investigated the post-thaw cell viability, morphology, proliferation capacity, basic stem cell characteristics, in vitro differentiation potential and apoptosis-related gene expression profile before and after cryopreservation. RESULTS: The cryoprotectant 10% DMSO has shown higher post-thaw cell viability of 81.2+/-0.58% whereas 10% PVP and cocktail solution have shown 62.87+/-0.35% and 72.2+/-0.23%, respectively at 0 h immediately thawing. The cell viability was further reduced in all the cryopreserved groups at 24 h later post-thaw culture. Further, the complete elimination of FBS in cryoprotectants has resulted in drastic reduction in cell viability. Cryopreservation did not alter the basic stem cell characteristics, plasticity and multipotency except proliferation rate. The expression of pro-apoptotic BAX and p53 genes were higher whilst p21 was lower in all the cryopreserved groups when compare to the control group of WJMSCs. CONCLUSION: Although 10% DMSO has shown higher post-thaw cell viability compare to 10% PVP and cocktail solution, the present study indicates the feasibility of developing a well-defined DMSO free cryosolution which can improve storage and future broad range applications of WJMSCs in regenerative medicine without losing their basic stem cell characteristics.
Apoptosis ; Cell Survival ; Cryopreservation* ; Dimethyl Sulfoxide ; Ethylene Glycol ; Freezing* ; Genes, p53 ; Glucose ; Humans* ; Mesenchymal Stromal Cells* ; Plastics ; Povidone ; Regenerative Medicine ; Stem Cells ; Sucrose ; Transcriptome ; Wharton Jelly

Bilateral sagittal split ramus osteotomy is considered a standard technique in mandibular orthognathic surgeries to reduce unexpected bilateral stress in the temporomandibular joints. Unilateral sagittal split ramus osteotomy (USSO) was recently introduced to correct facial asymmetry caused by asymmetric mandibular prognathism and has shown favorable outcomes. If unilateral surgery could guarantee long-term postoperative stability as well as favorable results, operation time and the incidence of postoperative complications could be reduced compared to those in bilateral surgery. This report highlights three consecutive cases with long-term follow-up in which USSO was used to correct asymmetric mandibular prognathism. Long-term postoperative changes in the condylar contour and ramus and condylar head length were analyzed using routine radiography and computed tomography. In addition, prior USSO studies were reviewed to outline clear criteria for applying this technique. In conclusion, patients showing functional-type asymmetry with predicted unilateral mandibular movement of less than 7 mm can be considered suitable candidates for USSO-based correction of asymmetric mandibular prognathism with or without maxillary arch surgeries.
Facial Asymmetry* ; Follow-Up Studies ; Head ; Humans ; Incidence ; Orthognathic Surgery ; Osteotomy, Sagittal Split Ramus* ; Postoperative Complications ; Prognathism ; Radiography ; Temporomandibular Joint

Bone and Bones ; Carbocyanines ; Cell Culture Techniques ; Coculture Techniques ; Collagen ; Collagenases ; Drug Combinations ; Durapatite ; Endothelial Cells ; Laminin ; Lipoproteins ; Magnetics ; Magnets ; Mandible ; Mesenchymal Stromal Cells ; Molar, Third ; Nylons ; Osteoblasts ; Osteogenesis ; Perchloric Acid ; Periosteum ; Proteoglycans

A conference was convened by the Korean Diabetes Association and the Korean Endocrine Society on September 7, 2009 to discuss and organize the results of research on intensive glucose control for the prevention of cardiovascular disease in patients with type 2 diabetes. Professor Kyung Soo Park led the conference, and Professors Kwang Won Kim and Ho Young Son acted as chairmen. Professors Doo Man Kim, Tae Sun Park, and Bong Soo Cha reported on intensive glucose control and diabetic complications, including the UK Prospective Diabetes Study (UKPDS), Diabetes Control and Complication Trial (DCCT) research results, the recently published Action to Control Cardiovascular Risk in Diabetes (ACCORD), Action in Diabetes and Vascular Disease: Preterax and Diamicron Modified Release Controlled Evaluation (ADVANCE), and Veterans Affairs Diabetes Trial (VADT) research, as well as meta-analyses. Professor Jeong-Taek Woo reported on the manuscript written by the committee for the Korean Diabetes Association which dealt with the treatment of diabetes mellitus. Professors Kyung Soo Ko, Joong Yeol Park, Hyun Shik Son, Moon-Kyu Lee, Dong-Won Byun, and Yoon-Sok Chung participated in the discussion and collected information for the manuscript from all of the participants. The aim of the debate was to determine how to establish target goals for intensive glucose control and how to individualize those goals. The participants concluded that there was no need to modify the recommendation of maintaining an HbA1c under 6.5%, the current blood glucose treatment goal that is recommended by the Korean Diabetes Association. In addition, individual target goals for glucose control were recommended depending on the situation of each patient. We report on the consensus statement from the meeting.
Blood Glucose ; Cardiovascular Diseases ; Consensus ; Diabetes Complications ; Diabetes Mellitus ; Drug Combinations ; Gliclazide ; Glucose ; Humans ; Indapamide ; Perindopril ; Solar System ; Veterans

A conference was convened by the Korean Diabetes Association and the Korean Endocrine Society on September 7, 2009 to discuss and organize the results of research on intensive glucose control for the prevention of cardiovascular disease in patients with type 2 diabetes. Professor Kyung Soo Park led the conference, and Professors Kwang Won Kim and Ho Young Son acted as chairmen. Professors Doo Man Kim, Tae Sun Park, and Bong Soo Cha reported on intensive glucose control and diabetic complications, including the UK Prospective Diabetes Study (UKPDS), Diabetes Control and Complication Trial (DCCT) research results, the recently published Action to Control Cardiovascular Risk in Diabetes (ACCORD), Action in Diabetes and Vascular Disease: Preterax and Diamicron Modified Release Controlled Evaluation (ADVANCE), and Veterans Affairs Diabetes Trial (VADT) research, as well as meta-analyses. Professor Jeong-Taek Woo reported on the manuscript written by the committee for the Korean Diabetes Association which dealt with the treatment of diabetes mellitus. Professors Kyung Soo Ko, Joong Yeol Park, Hyun Shik Son, Moon-Kyu Lee, Dong-Won Byun, and Yoon-Sok Chung participated in the discussion and collected information for the manuscript from all of the participants. The aim of the debate was to determine how to establish target goals for intensive glucose control and how to individualize those goals. The participants concluded that there was no need to modify the recommendation of maintaining an HbA1c under 6.5%, the current blood glucose treatment goal that is recommended by the Korean Diabetes Association. In addition, individual target goals for glucose control were recommended depending on the situation of each patient. We report on the consensus statement from the meeting.
Blood Glucose ; Cardiovascular Diseases ; Consensus ; Diabetes Complications ; Diabetes Mellitus ; Drug Combinations ; Gliclazide ; Glucose ; Humans ; Indapamide ; Perindopril ; Solar System ; Veterans

INTRODUCTION: Skeletal homeostasis is normally maintained by the stability between bone formation by osteoblasts and bone resorption by osteoclasts. However, the correlation between the inflammatory reaction and osteoblastic differentiation of cultured osteoprogenitor cells has not been fully investigated. This study examined the effects of inflammatory cytokines on the osteoblastic differentiation of cultured human periosteal-derived cells. MATERIALS AND METHODS: Periosteal-derived cells were obtained from the mandibular periosteum and introduced into the cell culture. After passage 3, the periosteal-derived cells were further cultured in an osteogenic induction Dulbecco's modified Eagle's medium (DMEM) medium containing dexamethasone, ascorbic acid, and beta-glycerophosphate. In this culture medium, tumor necrosis factor (TNF)-alpha with different concentrations (0.1, 1, and 10 ng/mL) or interleukin (IL)-1beta with different concentrations (0.01, 0.1, and 1 ng/mL) were added. RESULTS: Both TNF-alpha and IL-1beta stimulated alkaline phosphatase (ALP) expression in the periosteal-derived cells. TNF-alpha and IL-1beta increased the level of ALP expression in a dose-dependent manner. Both TNF-alpha and IL-1beta also increased the level of alizarin red S staining in a dose-dependent manner during osteoblastic differentiation of cultured human periosteal-derived cells. CONCLUSION: These results suggest that inflammatory cytokines TNF-alpha and IL-1beta can stimulate the osteoblastic activity of cultured human periosteal-derived cells.
Alkaline Phosphatase ; Anthraquinones ; Ascorbic Acid ; Bone Resorption ; Cell Culture Techniques ; Cytokines ; Dexamethasone ; Durapatite ; Glycerophosphates ; Homeostasis ; Humans ; Interleukins ; Osteoblasts ; Osteoclasts ; Osteogenesis ; Periosteum ; Tumor Necrosis Factor-alpha

INTRODUCTION: The first aim of this study was to isolate the dental tissue-derived stem cells from the dental follicle (DF), dental pulp (DP), and root apical papilla (RAP) of the extracted wisdom teeth. Second was to evaluate their characterization with the expressions of transcription factors and cell surface markers. Finally, their ability of the in vitro multi-lineage differentiations into osteogenic and adipogenic cells were compared, respectively. MATERIALS AND METHODS: Dental tissues, including dental follicle, dental pulp, and root apical papilla, were separated in the extracted wisdom teeth. These three dental tissues were cultured in Dulbecco's modified Eagle's medium (DMEM) with supplements, respectively. After passage 3, the homogeneous shaped dental tissue-derived cells were analyzed the expression of transcription factors (Oct-4, Nanog and Sox-2) and cell surface markers (CD44, CD90 and CD105) with reverse transcription polymerase chain reaction (RT-PCR) and fluorescence-activated cell sorting (FACS) analysis. In order to evaluate in vitro multi-lineage differentiations, the culture media were changed to the osteogenic and adipogenic induction mediums when the dental tissue-derived cells reached to passage 3. The characteristics of these three dental tissue-derived cells were compared with immunohistochemistry. RESULTS: During primary culture, heterogenous and colony formatted dental tissue-derived cells were observed in the culture plates. After passage 2 or 3, homogenous spindle-like cells were observed in all culture plates. Transcription factors and mesenchymal stem cell markers were positively observed in all three types of dental tissue-derived cells. However, the quantity of expressed transcription factors was most large in RAP-derived cells. In all three types of dental tissue-derived cells, osteogenic and adipogenic differentiations were observed after treatment of specific induction media. In vitro adipogenic differentiation was similar among these three types of cells. In vitro osteogenic differentiation was most strongly and frequently observed in the RAP-derived cells, whereas rarely osteogenic differentiation was observed in the DP-derived cells. CONCLUSION: These findings suggest that three types of human dental tissue-derived cells from extracted wisdom teeth were multipotent mesenchymal stem cells, have the properties of multi-lineage differentiations. Especially, stem cells from root apical papilla (SCAP) have much advantage in osteogenic differentiation, whereas dental follicle cells (DFCs) have a characteristic of easy adipogenic differentiation.
Culture Media ; Dental Pulp ; Dental Sac ; Durapatite ; Flow Cytometry ; Humans ; Imidazoles ; Immunohistochemistry ; Mesenchymal Stromal Cells ; Molar, Third ; Nitro Compounds ; Polymerase Chain Reaction ; Reverse Transcription ; Stem Cells ; Transcription Factors

Alkaline Phosphatase ; Ascorbic Acid ; Calcium ; Cell Culture Techniques ; Cell Proliferation ; Dexamethasone ; Durapatite ; Humans ; Mandible ; Molar, Third ; Osteoblasts ; Osteocalcin ; Phenotype ; RNA, Messenger ; Strontium

Adipose Tissue ; Adult ; Ascorbic Acid ; Bone Development ; Bone Marrow ; Cell Culture Techniques ; Cell Separation ; Collagen ; Collagenases ; Drug Combinations ; Durapatite ; Endothelial Cells ; Epidermal Growth Factor ; Flow Cytometry ; Heparin ; Humans ; Hydrocortisone ; Laminin ; Magnetics ; Magnets ; Mesenchymal Stromal Cells ; Microspheres ; Nylons ; Osteoblasts ; Osteotomy, Sagittal Split Ramus ; Prognathism ; Proteoglycans ; Skin ; Stem Cells ; Vascular Endothelial Growth Factor A

Collagen ; Drug Combinations ; Durapatite ; Endothelial Cells ; Humans ; Laminin ; Proteoglycans