Jong-Ryoul Kim

In Vitro Osteogenic Differentiation of Cultured Human Dental Papilla-Derived Cells

PURPOSE This study examined the osteogenic phenotypes and mineralization of cultured human dental papilla-derived cells. MATERIALS AND METHODS Dental papillae were harvested from mandibles during surgical extraction of lower impacted third molars from 3 patients aged 13 to 15 years. The dental papilla-derived cells were introduced into the cell culture. After passage 3, the dental papilla-derived cells were further cultured for 42 days in an osteogenic inductive culture medium containing dexamethasone, ascorbic acid, and beta-glycerophosphate. We examined the histochemical detection of alkaline phosphatase (ALP), the reverse transcriptase-polymerase chain reaction (RT-PCR) analysis for ALP and osteocalcin, and von Kossa staining in the dental papilla-derived cells. RESULTS It was observed that ALP was strongly expressed in the earlier stage of osteoblastic differentiation, whereas osteocalcin was mainly expressed and secreted into the medium at the later stage. Von Kossa-positive mineralization nodules were first observed on day 14, which increased in number during the entire culture period. CONCLUSIONS These results suggest that dental papilla-derived cell have osteogenic potential and could be used as an additional source of cells for bone tissue engineering.

Tissue-engineered bone formation using periosteal-derived cells and polydioxanone/pluronic F127 scaffold with pre-seeded adipose tissue-derived CD146 positive endothelial-like cells.

The aim of this study was to generate tissue-engineered bone formation using periosteal-derived cells seeded into a polydioxanone/pluronic F127 (PDO/Pluronic F127) scaffold with adipose tissue-derived CD146 positive endothelial-like cells. Considering the hematopoietic and mesenchymal phenotypes of adipose tissue-derived cells cultured in EBM-2 medium, CD146 positive adipose tissue-derived cells was sorted to purify more endothelial cells in characterization. These sorted cells were referred to as adipose tissue-derived CD146 positive endothelial-like cells. Periosteum is a good source of osteogenic cells for tissue-engineered bone formation. Periosteal-derived cells were found to have good osteogenic capacity in a PDO/Pluronic F127 scaffold, which could provide a suitable environment for the osteoblastic differentiation of these cells. Through the investigation of capillary-like tube formation on matrigel and the cellular proliferation of adipose tissue-derived CD146 positive endothelial-like cells cultured in different media conditions, we examined these cells could be cultured in EBM-2 with osteogenic induction factors. We also observed that the osteogenic activity of periosteal-derived cells could be good in EBM-2 with osteogenic induction factors, in the early period of culture. The experimental results obtained in the miniature pig model suggest that tissue-engineered bone formation using periosteal-derived cells and PDO/Pluronic F127 scaffold with pre-seeded adipose tissue-derived CD146 positive endothelial-like cells can be used to restore the bony defects of the maxillofacial region when used in clinics.

Three-dimensional analysis of midfacial soft tissue changes according to maxillary superior movement after horizontal osteotomy of the maxilla.

Three-dimensional diagnostic and treatment planning is a promising means of improving orthognathic surgical results from the standpoint of facial aesthetics. In such planning, cone-beam computed tomography is a useful tool. The purpose of the present study was to evaluate three-dimensional soft tissue changes in the midfacial region, with specific reference to post-Le Fort I osteotomy maxillary superior movement. Twenty-two patients underwent both Le Fort I osteotomy superior impaction and bilateral sagittal split ramus osteotomy of the mandible (ie, double-jaw surgery). Reference planes and 27 × 10 grids were used in evaluating the midfacial soft tissue areas. The extent of soft tissue change before and after surgery was calculated and analyzed. The results showed no statistical difference between the male and female subjects (P > 0.05). For both the male and the female patients after double-jaw surgery, the soft tissue in the triangular area, which includes both the nasolabial grooves and the upper lip, moved in the anterior direction and maxillary superiorly. It is essential that clinicians concerned about the management of soft tissue and the quality of treatment outcomes understand the pattern of soft tissue change after double-jaw surgery.

Vascular endothelial growth factor stimulates osteoblastic differentiation of cultured human periosteal-derived cells expressing vascular endothelial growth factor receptors

Angiogenesis plays an important role in bone development and postnatal bone fracture repair. Vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptors (VEGFRs) are primarily involved in angiogenesis. This study investigated the expression of VEGF isoforms, VEGFR-1, and VEGFR-2 during the osteoblastic differentiation of cultured human periosteal-derived cells. In addition, the effect of exogenous VEGF on the osteoblastic differentiation of cultured human periosteal-derived cells was also examined. The expression of the VEGF isoforms (VEGF121, VEGF165, VEGF189, and VEGF206), VEGFR-1, and VEGFR-2 was observed in the periosteal-derived cells. Administration of KRN633, a VEGFR-1 and VEGFR-2 inhibitor, decreased the alkaline phosphatase (ALP) activity during the osteoblastic differentiation of cultured human periosteal-derived cells. However, the administration of VEGFR2 Kinase Inhibitor IV, a VEGFR-2 inhibitor, did not affect the ALP activity. The addition of recombinant human VEGF165 elevated the ALP activity and increased the calcium content in the periosteal-derived cells. Treating the periosteal-derived cells with recombinant human VEGF165 resulted in an increase in Runx2 transactivation in the periosteal-derived cells. These results suggest that exogenous VEGF stimulates the osteoblastic differentiation of cultured human periosteal-derived cells and VEGF might act as an autocrine growth factor for the osteoblastic differentiation of cultured human periosteal-derived cells.

Vascular endothelial growth factor expression in cultured periosteal-derived cells.

The purpose of this study was to examine the expression of vascular endothelial growth factor (VEGF) during osteoblastic differentiation of cultured human periosteal-derived cells. Periosteal tissues were obtained from mandible during surgical extraction of lower impacted third molars. Periosteal-derived cells were introduced into cell culture. After passage 3, the periosteal-derived cells were further cultured for 42 days in an osteogenic-inductive culture medium containing dexamethasone, ascorbic acid, and beta-glycerophosphate. The alkaline phosphatase activity in the cultured periosteal-derived cells increased rapidly up to day 14, followed by decrease in activity. The Runx2 protein was expressed at day 7 and day 14, and its expression was not observed thereafter. Both VEGF(165) and VEGF(121) were expressed strongly at days 35 and 42 of culture, particularly during the later stages of differentiation. Alizarin red S-positive nodules were first observed on day 14 and then increased in number during the entire culture period. Osteocalcin and VEGF were first detected in the culture medium on day 14, and their levels increased thereafter in a time-dependent manner. These results suggest that VEGF secretion from cultured human periosteal-derived cells increases along with the mineralization process of the extracellular matrix.

New bimaxillary orthognathic surgery planning and model surgery based on the concept of six degrees of freedom

The aim of this paper was to propose a new method of bimaxillary orthognathic surgery planning and model surgery based on the concept of 6 degrees of freedom (DOF). A 22-year-old man with Class III malocclusion was referred to our clinic with complaints of facial deformity and chewing difficulty. To correct a prognathic mandible, facial asymmetry, flat occlusal plane angle, labioversion of the maxillary central incisors, and concavity of the facial profile, bimaxillary orthognathic surgery was planned. After preoperative orthodontic treatment, surgical planning based on the concept of 6 DOF was performed on a surgical treatment objective drawing, and a Jeons model surgery chart (JMSC) was prepared. Model surgery was performed with Jeons orthognathic surgery simulator (JOSS) using the JMSC, and an interim wafer was fabricated. Le Fort I osteotomy, bilateral sagittal split ramus osteotomy, and malar augmentation were performed. The patient received lateral cephalometric and posteroanterior cephalometric analysis in postretention for 1 year. The follow-up results were determined to be satisfactory, and skeletal relapse did not occur after 1.5 years of surgery. When maxillary and mandibular models are considered as rigid bodies, and their state of motion is described in a quantitative manner based on 6 DOF, sharing of exact information on locational movement in 3-dimensional space is possible. The use of JMSC and JOSS will actualize accurate communication and performance of model surgery among clinicians based on objective measurements.

Low-level laser therapy affects osseointegration in titanium implants: resonance frequency, removal torque, and histomorphometric analysis in rabbits.

Objectives The purpose of this study was to investigate the effects of low-level laser therapy (LLLT) with a diode gallium-aluminum-arsenide (Ga-Al-As) low-level laser device on the healing and attachment of titanium implants in bone. Materials and Methods Thirteen New Zealand white male rabbits weighing 3.0±0.5 kg were used for this study. Dental titanium implants (3.75 mm in diameter and 8.5 mm in length, US II RBM plus fixture; Osstem, Seoul, Korea) were implanted into both femurs of each rabbit. The rabbits were randomly divided into a LLLT group and a control group. The LLLT was initiated immediately after surgery and then repeated daily for 7 consecutive days in the LLLT group. Six weeks and 12 weeks after implantation, we evaluated and compared the osseointegration of the LLLT group and control group, using histomorphometric analysis, removal torque testing, and resonance frequency analysis (RFA). The results were statistically significant when the level of probability was 0.05 or less based on a non-parametric Mann-Whitney U-test. Results The implant survival rate was about 96%. Histologically and histomorphometrically, we observed that the titanium implants were more strongly attached in LLLT group than in control group. However, there was no significant difference between the LLLT group and control group in removal torque or RFA. Conclusion Histologically, LLLT might promote cell-level osseointegration of titanium implants, but there was no statistically significant effects.

Generation of osteogenic construct using periosteal‐derived osteoblasts and polydioxanone/pluronic F127 scaffold with periosteal‐derived CD146 positive endothelial‐like cells

The purpose of this study was to generate tissue-engineered bone using human periosteal-derived osteoblasts (PO) and polydioxanone/pluronic F127 (PDO/pluronic F127) scaffold with preseeded human periosteal-derived CD146 positive endothelial-like cells (PE). PE were purified from the periosteal cell population by cell sorting. One of the important factors to consider in generating tissue-engineered bone using osteoprecursor and endothelial cells and a specific scaffold is whether the function of osteoprecursor and endothelial cells can be maintained in originally different culture medium conditions. After human PE were preseeded into PDO/pluronic F127 scaffold and cultured in endothelial cell basal medium-2 for 7 days, human PO were seeded into the PDO/pluronic F127 scaffold with PE, and then, this cell-scaffold construct was cultured in endothelial cell basal medium-2 with osteogenic induction factors, including ascorbic acid, dexamethasone, and β-glycerophosphate, for a further 7 days. Then, this 2-week cultured construct was grafted into the mandibular defect of miniature pig. Twelve weeks after implantation, the animal was sacrificed. Clinical examination revealed that newly formed bone was seen more clearly in the defect with human PO and PDO/pluronic F127 scaffold with preseeded human PE. The experimental results suggest that tissue-engineered bone formation using human PO and PDO/pluronic F127 scaffold with preseeded human PE can be used to restore skeletal integrity to various bony defects when used in clinics.

Stability of unilateral sagittal split ramus osteotomy for correction of facial asymmetry: long-term case series and literature review

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.