Effect of CD146+ SHED on bone regeneration in a mouse calvaria defect model.

OBJECTIVE: Stem cells from human exfoliated deciduous teeth (SHED) have bone regeneration ability and potential therapeutic applications. CD146, a cell adhesion protein expressed by vascular endothelial cells, is involved in osteoblastic differentiation of stem cells. The effect of CD146 on SHED-mediated bone regeneration in vivo remains unknown. We aimed to establish efficient conditions for SHED transplantation. MATERIALS AND METHODS: SHED were isolated from the pulp of an extracted deciduous tooth and cultured; CD146-positive (CD146+ ) and CD146-negative (CD146- ) populations were sorted. Heterogeneous populations of SHED and CD146+ and CD146- cells were transplanted into bone defects generated in the skulls of immunodeficient mice. Micro-computed tomography was performed immediately and 4 and 8 weeks later. Histological and immunohistochemical assessments were performed 8 weeks later. RESULTS: Bone regeneration was observed upon transplantation with CD146+ and heterogeneous populations of SHED, with significantly higher bone regeneration observed with CD146+ cells. Bone regeneration was higher in the CD146- group than in the control group, but significantly lower than that in the other transplant groups at 4 and 8 weeks. Histological and immunohistochemical assessments revealed that CD146+ cells promoted bone regeneration and angiogenesis. CONCLUSION: Transplantation of CD146+ SHED into bone defects may be useful for bone regeneration.

Bone Differentiation Ability of CD146-Positive Stem Cells from Human Exfoliated Deciduous Teeth.

Regenerative therapy for tissues by mesenchymal stem cell (MSCs) transplantation has received much attention. The cluster of differentiation (CD)146 marker, a surface-antigen of stem cells, is crucial for angiogenic and osseous differentiation abilities. Bone regeneration is accelerated by the transplantation of CD146-positive deciduous dental pulp-derived mesenchymal stem cells contained in stem cells from human exfoliated deciduous teeth (SHED) into a living donor. However, the role of CD146 in SHED remains unclear. This study aimed to compare the effects of CD146 on cell proliferative and substrate metabolic abilities in a population of SHED. SHED was isolated from deciduous teeth, and flow cytometry was used to analyze the expression of MSCs markers. Cell sorting was performed to recover the CD146-positive cell population (CD146+) and CD146-negative cell population (CD146-). CD146 + SHED without cell sorting and CD146-SHED were examined and compared among three groups. To investigate the effect of CD146 on cell proliferation ability, an analysis of cell proliferation ability was performed using BrdU assay and MTS assay. The bone differentiation ability was evaluated using an alkaline phosphatase (ALP) stain after inducing bone differentiation, and the quality of ALP protein expressed was examined. We also performed Alizarin red staining and evaluated the calcified deposits. The gene expression of ALP, bone morphogenetic protein-2 (BMP-2), and osteocalcin (OCN) was analyzed using a real-time polymerase chain reaction. There was no significant difference in cell proliferation among the three groups. The expression of ALP stain, Alizarin red stain, ALP, BMP-2, and OCN was the highest in the CD146+ group. CD146 + SHED had higher osteogenic differentiation potential compared with SHED and CD146-SHED. CD146 contained in SHED may be a valuable population of cells for bone regeneration therapy.

Effect of high-frequency near-infrared diode laser irradiation on periodontal tissues during experimental tooth movement in rats.

BACKGROUND AND OBJECTIVES: Tooth movement during orthodontic treatment is associated with bone neoplasticity and bone resorption on the tension and pressure sides. Previous clinical studies have suggested that low-power laser irradiation can accelerate tooth movement during orthodontic treatment, although the underlying mechanism remains unclear. In this study, we used a high-frequency near-infrared diode laser that generates less heat and examined the histologic changes in periodontal tissue during experimental tooth movement with laser irradiation. METHODS: A nickel-titanium closed coil was mounted between the maxillary left side first molar and incisor of rats to model experimental tooth movement. The laser-irradiation and the control groups were set, and the amount of movement of the first molar on 7th and 14th days after the start of pulling of the first molar tooth on the maxillary left was measured by three-dimensional analysis of µCT. After tooth movement, tissue samples from the mesial and tension sides were collected, and successive horizontal sections were prepared and examined using hematoxylin-eosin and TRAP staining and immunohistochemical staining for RANKL, OPG, ALP, and proliferating cell nuclear antigen (PCNA). Changes in tissue temperature following laser irradiation were also examined. RESULTS: Laser irradiation significantly increased tooth movement compared with non-irradiated controls. Histologic staining of the pressure-side mesial root in laser-irradiated rats revealed enhanced RANKL expression and increased numbers of TRAP-positive cells compared with controls. By contrast, on the tension side, laser irradiation led to increased expression of ALP and PCNA. These data indicate that high-frequency near-infrared diode laser irradiation on the pressure side upregulates RANKL expression and accelerates osteoclast differentiation, facilitating bone resorption, whereas bone formation is induced on the tension side. CONCLUSION: This study demonstrates that high-frequency near-infrared diode laser irradiation of periodontal tissue leads to metabolic activation, which ultimately increases the rate of tooth movement. Lasers Surg. Med. © 2018 Wiley Periodicals, Inc.

Accuracy of a three-dimensional dentition model digitized from an interocclusal record using a non-contact surface scanner.

OBJECTIVES: For orthodontic treatment, it is important to assess the dental morphology, as well as the position and inclination of teeth. The aim of this article was to develop an efficient and accurate method for the three-dimensional (3D) imaging of the maxillary and mandibular dental morphology by measuring interocclusal records using an optical scanner. MATERIALS AND METHODS: The occlusal and incisal morphology of participants was registered in the intercuspal position using a hydrophilic vinyl polysiloxane and digitized into 3D models using an optical scanner. Impressions were made of the maxilla and mandible in alginate materials in order to fabricate plaster models and created into 3D models using the optical scanner based on the principal triangulation method. The occlusal and incisal areas of the interocclusal records were retained. The buccal and lingual areas were added to these regions entirely by the 3D model of the plaster model. The accuracy of this method was evaluated for each tooth, with the dental cast 3D models used as controls. RESULTS: The 3D model created from the interocclusal record and the plaster model of the dental morphology was analysed in 3D software. The difference between the controls and the 3D models digitized from the interocclusal records was 0.068±0.048mm, demonstrating the accuracy of this method. LIMITATIONS: The presence of severe crowding may compromise the ability to separate each tooth and digitize the dental morphology. CONCLUSIONS: The digitization method in this study provides sufficient accuracy to visualize the dental morphology, as well as the position and inclination of these teeth.

Maxillofacial Morphology as a Predictive Factor for Caries Risk in Orthodontic Patients: A Cross-Sectional Study.

This cross-sectional study aimed to explore the correlation between maxillofacial morphology and caries risk, assessed using salivary tests, in orthodontic patients. Despite enhancing the oral health-related quality of life, orthodontic treatment may adversely affect oral hygiene and increase caries risk. This study included 1071 patients all of whom underwent orthodontic examinations and salivary tests before starting orthodontic treatment at a hospital. Salivary tests were performed to assess the secretion rate, pH, buffering capacity, and counts of cariogenic bacteria. The maxillofacial morphology was evaluated using cephalometric X-rays and dental models. Statistical analyses revealed significant correlations among salivary characteristics, bacterial scores, and maxillofacial morphology. Notably, the facial angle and Y-axis values were associated with salivary secretion (p < 0.001), pH (p < 0.001), buffering capacity (p < 0.05), and cariogenic bacterial scores (p < 0.01), respectably. In conclusion, assessing the maxillofacial morphology before orthodontic treatment may aid in predicting the risk of bacterial oral diseases, offering valuable insights into personalized preventive measures. These findings underscore the potential for comprehensive evaluations to enhance caries risk assessment in orthodontic patients.

Stem cells derived from human exfoliated deciduous teeth-based media in a rat root resorption model.

OBJECTIVE: Root resorption may occur during orthodontic treatment. Herein, we investigated the effect of a culture supernatant of stem cells derived from human exfoliated deciduous teeth on root resorption. DESIGN: Twelve 8-week-old male Sprague-Dawley rats were used, and their maxillary first molars were pulled with excessive orthodontic force to induce root resorption. On days 1 and 7 after traction initiation, stem cells derived from human exfoliated deciduous teeth and alpha minimum essential medium (control group) were administered. After 14 days, the maxillary bone was evaluated for tooth movement. The expression of osteoprotegerin, receptor activator of nuclear factor κB ligand, tumor necrosis factor α, interleukin 1ß, interleukin 6, and interleukin 17 was evaluated on the compression side and tension side. RESULTS: No significant difference in tooth movement was observed between the two groups. Root resorption decreased in the group administered the culture supernatant compared with in the control. Immunohistochemical staining revealed increased osteoprotegerin expression and decreased receptor activators for nuclear factor κB ligand, tumor necrosis factor α, interleukin 1ß, interleukin 6, and interleukin 17 on the compression side and tension side. CONCLUSIONS: Administration of stem cells derived from human exfoliated deciduous teeth affected the expression of osteoprotegerin, receptor activator of nuclear factor κB ligand, tumor necrosis factor α, interleukin 1ß, interleukin 6 and interleukin 17; hence, these stem cells may inhibit root resorption by regulating their expression.

Surgical Orthodontic Treatment for Skeletal Maxillary Protrusion in Sturge-Weber Syndrome: A Case Report and Review of the Literature.

Sturge-Weber syndrome (SWS) is characterized by hemangiomas, glaucoma, and central nervous system disorders. Here, we report the case of a 15-year-old boy with SWS and upper-lip hypertrophy who underwent surgical orthodontic treatment for correction of a large overjet and deep overbite. In addition to the a large overjet and deep overbite, interdental spacing was observed in both the arches. The mandible was retrognathic and deviated to the right side. No maxillary occlusal canting or temporomandibular joint symptoms were observed. The patient was diagnosed with skeletal maxillary protrusion with spaced dentition and mandibular deviation to the right due to SWS. After presurgical orthodontic treatment using a multibracket appliance, we performed a sagittal split ramus osteotomy (SSRO) alone due to the presence of a hemangioma around the maxilla. No abnormal bleeding or cerebral hemorrhage due to increased blood pressure was observed during the SSRO. Postoperatively, the maxillary and mandibular arches were well-aligned, the deep overbite and excessive overjet improved, and bilateral angle class I molar and canine relationships were established. Furthermore, mandibular deviation improved, and the midlines of both arches approximately coincided with the facial midline. In conclusion, orthognathic surgery is feasible in patients with SWS after carefully evaluating the sites and sizes of the hemangiomas.

RadImageNet and ImageNet as Datasets for Transfer Learning in the Assessment of Dental Radiographs: A Comparative Study.

Transfer learning (TL) is an alternative approach to the full training of deep learning (DL) models from scratch and can transfer knowledge gained from large-scale data to solve different problems. ImageNet, which is a publicly available large-scale dataset, is a commonly used dataset for TL-based image analysis; many studies have applied pre-trained models from ImageNet to clinical prediction tasks and have reported promising results. However, some have questioned the effectiveness of using ImageNet, which consists solely of natural images, for medical image analysis. The aim of this study was to evaluate whether pre-trained models using RadImageNet, which is a large-scale medical image dataset, could achieve superior performance in classification tasks in dental imaging modalities compared with ImageNet pre-trained models. To evaluate the classification performance of RadImageNet and ImageNet pre-trained models for TL, two dental imaging datasets were used. The tasks were (1) classifying the presence or absence of supernumerary teeth from a dataset of panoramic radiographs and (2) classifying sex from a dataset of lateral cephalometric radiographs. Performance was evaluated by comparing the area under the curve (AUC). On the panoramic radiograph dataset, the RadImageNet models gave average AUCs of 0.68 ± 0.15 (p < 0.01), and the ImageNet models had values of 0.74 ± 0.19. In contrast, on the lateral cephalometric dataset, the RadImageNet models demonstrated average AUCs of 0.76 ± 0.09, and the ImageNet models achieved values of 0.75 ± 0.17. The difference in performance between RadImageNet and ImageNet models in TL depends on the dental image dataset used.

Image preprocessing with contrast-limited adaptive histogram equalization improves the segmentation performance of deep learning for the articular disk of the temporomandibular joint on magnetic resonance images.

OBJECTIVES: The objective was to evaluate the robustness of deep learning (DL)-based encoder-decoder convolutional neural networks (ED-CNNs) for segmenting temporomandibular joint (TMJ) articular disks using data sets acquired from 2 different 3.0-T magnetic resonance imaging (MRI) scanners using original images and images subjected to contrast-limited adaptive histogram equalization (CLAHE). STUDY DESIGN: In total, 536 MR images from 49 individuals were examined. An expert orthodontist identified and manually segmented the disks in all images, which were then reviewed by another expert orthodontist and 2 expert oral and maxillofacial radiologists. These images were used to evaluate a DL-based semantic segmentation approach using an ED-CNN. Original and preprocessed CLAHE images were used to train and validate the models whose performances were compared. RESULTS: Original and CLAHE images acquired on 1 scanner had pixel values that were significantly darker and with lower contrast. The values of 3 metrics-the Dice similarity coefficient, sensitivity, and positive predictive value-were low when the original MR images were used for model training and validation. However, these metrics significantly improved when images were preprocessed with CLAHE. CONCLUSIONS: The robustness of the ED-CNN model trained on a dataset obtained from a single device is low but can be improved with CLAHE preprocessing. The proposed system provides promising results for a DL-based, fully automated segmentation method for TMJ articular disks on MRI.

Construction of orthodontic setup models on a computer.

INTRODUCTION: Orthodontic setup models are usually limited to the display of teeth, with no information about the roots. The purpose of this article is to present a method for visualizing the tooth roots in setup models by integrating information from cone-beam computed tomography and a laser scanner. The reproducibility of the integration was evaluated. METHODS: The records of 5 patients were used in this study. Three-dimensional digital models were generated from the dental casts. Tooth models were generated from the cone-beam computed tomography slices. The 3-dimensional models were superimposed on the crowns of the teeth in the tooth models and integrated. The integrated 3-dimensional tooth model and 3-dimensional setup model were registered. The reproducibility of the integration was evaluated for each tooth. Unpaired Student t tests were performed on the data between the anterior and posterior teeth, and between the right and left teeth. RESULTS: The discrepancy among the integrated 3-dimensional models at the final positions after we used this technique was 0.025 ± 0.007 mm. There was a significant difference in the distance between the anterior and posterior teeth (P <0.05). However, the average distances between the anterior and posterior teeth were small: 0.023 ± 0.007 and 0.028 ± 0.007 mm, respectively. No significant difference was found between the right and left teeth (P = 0.831). CONCLUSIONS: The methods presented in this study provide a reproducible visualization of tooth roots in virtual setup models by registering accurate crown models to cone-beam computed tomography scans.

Impact of Maximum Tongue Pressure in Patients with Jaw Deformities Who Underwent Orthognathic Surgery.

Malocclusion and morphological abnormalities of the jawbone often affect the stomatognathic function and long-term postoperative stability in patients with jaw deformities. There are few reports on the effect of maximum tongue pressure (MTP) in these patients. We investigated the relationship between the MTP and jawbone morphology and the effect of the MTP on surgery in 42 patients with jaw deformity who underwent surgical orthodontic treatment at Hiroshima University Hospital. The MTP was measured using a tongue pressure measurement device; the average value was considered as the MTP. Based on the MTP measured before surgery, patients were classified into the high- or the low-MTP group. The clinical findings and results of the cephalometric analysis were compared. Posterior movement of the mandible in the high-MTP group was significantly lower than that in the low-MTP group. The ANB angle, overjet, and overbite in the high-MTP group were significantly smaller than those in the low-MTP group. On the other hand, there was no difference between the two groups in the measured values, indicating a labial inclination of the anterior teeth (U1 to SN, U1 to FH, IMPA, and FMIA). MTP has been suggested to affect mandibular prognathism in patients with jaw deformities.

The Influence of 2-Methacryloyloxyethyl Phosphorylcholine Polymer Materials on Orthodontic Friction and Attachment of Oral Bacteria.

There is no clinical evidence of the usage of 2-methacryloyloxyethyl phosphorylcholine (MPC) polymers in dental practice. We performed in vitro studies to determine whether the application of an MPC coating to stainless steel orthodontic wires confers low-friction and antimicrobial properties to these wires. The friction test on MPC-coated wires was performed using a precision universal/tensile tester. MPC polymer was coated on a 50 × 50 mm stainless steel plate, and samples were assessed using an antimicrobial activity test. To verify the effect of MPC polymer-treated wires on experimental tooth movement models in vitro, examinations were performed on typodonts to determine the improvement in tooth movement efficiency. The polymer treatment wire groups demonstrated significantly enhanced tooth movement compared with the untreated wire groups, at both 50 g and 100 g traction forces. The results indicated that MPC coating inhibited the attachment of oral bacteria, such as Streptococcus mutans, on a stainless steel plate. Additionally, the coating seemed to improve the efficiency of tooth movement by reducing the occurrence of friction. The application of an MPC coating onto stainless steel wires, which are used as orthodontic materials, may reduce static friction and bacterial adherence to the oral cavity and improve tooth movement.

Landmark annotation and mandibular lateral deviation analysis of posteroanterior cephalograms using a convolutional neural network.

BACKGROUND/PURPOSE: Facial asymmetry is relatively common in the general population. Here, we propose a fully automated annotation system that supports analysis of mandibular deviation and detection of facial asymmetry in posteroanterior (PA) cephalograms by means of a deep learning-based convolutional neural network (CNN) algorithm. MATERIALS AND METHODS: In this retrospective study, 400 PA cephalograms were collected from the medical records of patients aged 4 years 2 months-80 years 3 months (mean age, 17 years 10 months; 255 female patients and 145 male patients). A deep CNN with two optimizers and a random forest algorithm were trained using 320 PA cephalograms; in these images, four PA landmarks were independently identified and manually annotated by two orthodontists. RESULTS: The CNN algorithms had a high coefficient of determination (R 2 ), compared with the random forest algorithm (CNN-stochastic gradient descent, R 2  = 0.715; CNN-Adam, R 2  = 0.700; random forest, R 2  = 0.486). Analysis of the best and worst performances of the algorithms for each landmark demonstrated that the right latero-orbital landmark was most difficult to detect accurately by using the CNN. Based on the annotated landmarks, reference lines were defined using an algorithm coded in Python. The CNN and random forest algorithms exhibited similar accuracy for the distance between the menton and vertical reference line. CONCLUSION: Our findings imply that the proposed deep CNN algorithm for detection of facial asymmetry may enable prompt assessment and reduce the effort involved in orthodontic diagnosis.

Surveillance of salivary properties of pre-orthodontic patients in relation to age and sex.

Saliva plays an important role in masticatory function and protection from dental caries. Although studies have been conducted on saliva properties, their results vary widely depending on population settings. Hence, this study was performed to evaluate the results of saliva properties in individuals who attended their first visit for orthodontic treatment. A total of 619 participants were included (387 females and 232 males; mean age: 14.6 years). We conducted oral examinations and saliva (stimulated) tests and evaluated the saliva flow rate, pH, and buffering capacity, along with bacterial culture. Saliva flow rate, pH, and buffering capacity were significantly higher in males than in females. However, the Streptococcus mutans score was significantly higher in females than in males even though oral hygiene was better in females. Significant positive correlations were found between age and saliva flow rate and S. mutans score. On the contrary, significant negative correlations were found between age and pH and buffering capacity. These results were similar to other studies where the target population was children or teenagers. Saliva properties of patients starting orthodontic treatment were almost the same as in populations of similar ages.

Baicalin inhibits root resorption during tooth movement in a rodent model.

OBJECTIVE: Baicalin mediates bone metabolism and has shown protective activity against periodontal tissue damage in a rat model of periodontitis. Therefore, we hypothesized that baicalin may inhibit the root resorption that occurs during orthodontic tooth movement and examined its effect on the histological changes in periodontal tissue that occur during tooth movement. METHODS: First molars of rats were subjected to traction using excessive orthodontic force to produce a root resorption model. Rats in the baicalin group received baicalin for 3 weeks during tooth movement, and the amount of first molar movement on day 21 after the initiation of traction was measured by three-dimensional micro-computed tomography analysis. After tooth movement, tissue samples from the mesial and tension sides were collected, and successive horizontal sections were prepared and examined using hematoxylin-eosin and tartrate-resistant acid phosphatase (TRAP) staining and immunohistochemical staining for the receptor activator of NF-kB ligand (RANKL) and osteoprotegerin (OPG). The severity of root resorption was also determined by histological analysis. RESULTS: There was no significant intergroup difference in tooth movement during the experimental exaggerated tooth movement. In comparison with the control group, the baicalin-treated group showed increased OPG expression, suppressed RANKL expression, and significantly fewer TRAP-positive cells in the first molars. The root resorption area was significantly smaller in the baicalin group. CONCLUSIONS: Treatment with baicalin prevented root resorption without preventing tooth movement. Baicalin may be useful for the management of root resorption during orthodontic treatment.

Effects of bFGF on the Modulation of Apoptosis in Gingival Fibroblasts with Different Host Ages.

The purpose of this study was to investigate the effects of basic fibroblast growth factor (bFGF) treatment on the proliferation and apoptosis of cultured gingival fibroblasts (GFs). Human GFs were isolated from the palatal gingival tissues of 16 healthy volunteers ranging in the age from 9 to 35 years old. Cultured GFs were subjected to the analyses for cell proliferation by ELISA assay, gene expression by RT-PCR analysis, and apoptosis potency by caspase-3 assay. The cell proliferation activity and gene expression of type-I collagen and caspase-3 activity were enhanced significantly by the treatment with bFGF in cultured GFs. Furthermore, the activity of caspase-3 in cultured GFs from young subjects was significantly higher than that in GFs from adults. It is shown that bFGF significantly enhances the gene expression of type-I collagen in cultured fibroblasts from human gingival tissues. It also demonstrated that bFGF modulates the apoptosis of periodontal fibroblasts, and the effect is higher in young subjects, indicating a significant role of bFGF in the prevention of scar formation during wound healing.

Differential effects of amelogenin on mineralization of cementoblasts and periodontal ligament cells.

BACKGROUND: Amelogenin is a major component of developing extracellular enamel matrix proteins and plays a crucial role during the formation of tooth enamel. In addition, amelogenins are suggested to exert biologic functions as signaling molecules through cell-surface receptors. The purpose of this study is to examine the effect of recombinant human full-length amelogenin (rh174) on the mineralization of human cementoblasts (HCEMs) and human periodontal ligament cells (HPDLs). METHODS: HCEMs, namely, a cell line immortalized by transfection of human telomerase reverse transcription gene, and HPDLs isolated from human first premolars were cultured and treated with 0 to 1,000 ng/mL rh174. The messenger ribonucleic acid (mRNA) levels of alkaline phosphatase (ALP), osteocalcin (OCN), and bone sialoprotein (BSP) were examined by real-time polymerase chain reaction analysis. The protein levels of OCN and BSP were examined by Western blot analysis. ALP activity and calcium deposition of cell cultures were also determined. Mineralization of cells was evaluated by red dye staining. RESULTS: The treatment of HCEMs with rh174 upregulated the ALP, OCN, and BSP mRNA levels. In addition, the protein levels of OCN and BSP, ALP activity, and calcium deposition were enhanced, resulting in enhanced mineralization. Conversely, there were no significant effects of rh174 on the mineralization of HPDLs. CONCLUSION: The present study shows that rh174 enhances mineralization accompanied by upregulation of mineralization markers in HCEMs, whereas it has no effect on that in HPDLs, suggesting different effects of amelogenin on PDL and cementum.

Amelogenin enhances the proliferation of cementoblast lineage cells.

BACKGROUND: It is well known that enamel matrix proteins play a crucial role in tooth root formation and amelogenesis. Because amelogenin is a major enamel matrix protein, it is assumed that amelogenin also affects the metabolism of cementum. However, the biologic functions of amelogenin in cementoblasts remain unclear. The purpose of this study is to examine the effect of recombinant human full-length amelogenin (rh174) on the proliferation of cultured human cementoblast-like (HCEM) and human periodontal ligament (HPDL) cells. METHODS: HCEM and HPDL cells were cultured and treated with 100 ng/mL rh174 in the presence or absence of an anti-cluster of differentiation (CD) 63 blocking antibody. Cell proliferation was evaluated using a cell proliferation enzyme-linked immunosorbent assay 5-bromo-2-deoxyuridine kit and quantification of the cell number by 3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2 H-tetrazolium-inner salt assay. The phosphorylation of extracellular signal-regulated kinases (ERK) 1/2 was measured by enzyme-linked immunosorbent assay and Western blot analysis. RESULTS: The proliferation of HCEM and HPDL cells was enhanced significantly (P <0.05) by treatment with rh174, and inhibited significantly (P <0.05) by the addition of anti-CD63 blocking antibody. In addition, the ratio of phosphorylated ERK1/2 to total ERK1/2 became significantly larger (P <0.05) by treatment with rh174, and was reduced significantly by the addition of anti-CD63 blocking antibody in both HCEM and HPDL cells. CONCLUSION: The results show that rh174 interacts with CD63, and rh174/CD63 interaction activates the ERK1/2 signaling pathway, enhancing the proliferation activities of HCEM and HPDL cells.