Wnt family regulates cell growth, differentiation and proliferation during embryonic development and also plays important roles in tumorigenesis. By the method of immunohistochemistry, a paralleled stydy was conducted to check Wnt5a and β-Catenin expression in OSCC formalin fixed-paraffin embedded tissues. The results demonstrated (1) Wnt5a and β-Catenin cytoplasmic expression appeared in most OSCC. (2) Statistical results showed positive correlation between the level of Wnt5a expression and OSCC differentiation (P<0.05) and inverse correlation between the level of β-Catenin cytoplasmic expression and OSCC differentiation (P<0.05). Cytoplasmic accumulation of β-Catenin protein also associated with lymph node metastasis (P<0.05), and loss of β-Catenin protein on cell membrane was correlative differentiation level of OSCC inversely (P<0.05). These data indicate Wnt/β-Catenin pathway associates with tumorigenesis, metastasis and prognosis of OSCC, and Wnt5a also associates with histological grade of OSCC.

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We focused attention of the calcification ability of coral cells. A section of coral was transplanted to the cavity of a tooth and observed microscopically for calcification after 7 days (LI group) and 14 days (LII group) of artificial irradiation. Observation after 7 days (NI group) and 14 days (NII group) without artificial irradiation was used as a control. Calcification was observed in the LI and LII groups, while it was not observed in the NI or NII group. The results suggest that coral contains single-cell xanthella which accelerate calcification by photosynthesis.

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Abnormal amplification of the epidermal growth factor receptor (EGFR) gene has been reported in various human tumors. We used competitive polymerase chain reaction (PCR) to study weather EGFR gene is amplified and the degree of amplification. We used in this study 17 cases of oral epithelial dysplasia (ED), 4 cases of carcinoma in Situ (CIS) and 20 cases of primary squamous cell carcinoma SCC. The extracted DNA was subjected to competitive PCR to amplify EGFR gene. Amplification of EGFR gene was observed in 3 cases (17%) of ED, 1 case of CIS and 4 cases (20%) of SCC. The degree of amplification was low in ED and CIS but extremely high in SCC. This suggests that amplification of EGFR occurs in early stage of oral SCC however high levels of EGFR plays a role in the progression to invasive carcinoma.

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Preferential alignment of biological apatite (BAp) c-axis and the related collagen (Col) fibril was proved to be a dominant parameter showing bone quality for understanding nano-scale microstructure and the related mechanical function in addition to bone mineral density (BMD). We clarified the correlations between in vivo stress distribution and the BAp/Col alignment and between the BAp/Col alignment and mechanical function, especially Young's modulus, in original intact and regenerative hard tissues. The mutual relationships are predicted to be closely related to the function of osteocyte which can sense the surrounding stress field. The BAp orientation was finally concluded to be one of the most important indices to evaluate in vivo stress distribution, nano-scale microstructure and the related mechanical function, regenerative process of the regenerated bone and progress of bone diseases.

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A novel web-like structured, biodegradable, hybrid sheet has been developed for bone tissue engineering by preparing knitted poly(DL-lactic-co-glycolic acid) (PLGA) sheets with collagen microsponges in their openings. The PLGA skeleton facilitated formation of the hybrid sheets into desired shapes, and the collagen microsponges in the pores of the PLGA sheet promoted cell adhesion and uniform cell distribution throughout the sheet. A large number of osteoblasts established from marrow stroma adhered to the scaffolds and generated the desired shaped bone in combination with this novel sheets. The web-like structured novel sheet show promise for use as a tool for custom-shaped bone regeneration in basic research on osteogenesis and for the development of therapeutic applications.

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From a subtracted cDNA library of human salivary gland a C77-91 gene totally unknown to date was named as Salvic that expresses 46 amino acids peptide (pI=9.45) possessing an antimicrobial activity on E. coli. Salvic is consisted of a typical hydrophobic amino acid rich domain in the N-terminus, a cluster of basic amino acids, carbohydrate attachment sites, a possible transglutaminase catalyzed crosslinking site, and multiple consensus sequences of phosphorylation site in the C-terminus. Western blot analysis using the monospecific antibody to the synthetic Salvic peptide showed strong interacting proteins in the extracts from submandibular gland and parotid saliva, and the immunohistochemical staining detected a strong positive reaction in the cytoplasmic secretory granules of interlobular ductal cells of salivary gland. The Salvic was also distributed in the human sebaceous gland and prostate. These data suggest that the identification of Salvic may add further understanding of greater role of salivary proteins providing innate immunity by protecting and stabilizing the mucosal epithelium in the maintaining homeostasis of oral mucosa.

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Our purpose in the present study was to establish a system for in vitro hepatic morphogenesis, consisting of not only hepatocytes but also cell lineages supporting hepatic differentiation, such as cardiomyocytes and endothelial cells, from murine ES cells. We succeeded in establishing a novel system of hepatic morphogenesis from murine ES cells based on naturally occurring embryological events.

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Adhesives for tooth bonding, implants, bone substitutes, bone cements, etc. are widely used for functional reconstruction of human hard tissues, such as bone and tooth tissue. The success of these treatments depends to a large extent on the properties of the resultant biomaterial-hard tissue interface. Thus, it is our belief that if the biomaterial surface can be nano-technologically controlled, more intense interaction of the biomaterial with the hard tissue can be achieved. In this paper, we report on three examples of nano-technological control of biomaterial-hard tissue interfaces, as there are the bonding of a so-called dental self-etch adhesive to tooth tissue, the integration of biological apatite in bone and the modification of the surface of Ti implants to promote cell attachment and growth.

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Human bone marrow mesenchymal stem cells (hMSCs), tissue-derived stem cells, have the ability to differentiate into adipocytes, osteoblasts and chondrocytes, leading to attempt to apply these stem cells for regeneration of bone and cartilage. To address pluripotency of hMSCs, we analyzed gene expression profiles of several immortalized hMSCs with DNA chips that nearly covers the whole human genes. We isolated some factors that possibly relate to pluripotency of these hMSCs.

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We studied the metallothionein (MT) induction by cadmium (Cd) in the dental pulp and bone in rats. In order to clarify the cell response to Cd in the hard tissue, the isoform-specific expression of MT mRNAs (MT-I and MT-II) was examined by reverse transcriptase-polymerase chain reaction (RT-PCR) method, and the localization of MT was confirmed by immunohistochemical studies. Both MT-I and MT-II mRNA levels were increased within 3 hours by Cd administration in both the dental pulp and bone. In the dental pulp, MT protein was localized in the specific cell type of odontoblasts (secretory odontoblasts and resting odontoblasts). In the bone, MT-positive cells were time-dependently increased, and the positive cells were mainly localized in osteocytes. The cell-specific induction of MT may be associated with Cd accumulation and Cd-induced the hard tissue injury in vivo.

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The present study was undertaken to investigate the effects of long term administration of valproate that is commonly used in clinics and co-administration of alfacalcidol or alendronate with valproate on bone metabolism in growing rats. Bone mineral densities and serum bone makers were determined. The samples obtained from tibial metaphysis were then prepared for histological observation. As the results, valproate decreased the bone mineral density in both metaphysis and diaphysis with dose dependent manner. The magnitude of the decrease was greater in the diaphysis than metaphysis. Decreased bone volume was observed in all of groups treated with valproate. Although the levels of serum calcium were not affected, valproate increased the activities of acid phosphatase and alkaline phosphatase in the serum. The above results showed that valproate could induce bone loss in growing rats. Possible reason of the decrease was due to bone loss. Combined administration of alfacalcidol or alendronete prevented the osteopnenia induced by valproate. Therefore, these drugs cloud have useful efficacy to prevent osteopenia induced by valproate.

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We investigated the effect of epiregulin (EPR) and EGF on the bone resorption using mice bones and cells. Both EPR and EGF enhanced bone resorption in ⁴⁵Ca-prelabeled parietal bone without affecting osteoclast differentiation. These growth factors also enhanced the pit formation by femur-derived osteoclastrich fraction without increasing the number of the osteoclasts in a NS-398 (a COX-2 inhibitor)- sensitive manner. Furthermore, mature osteoclasts, differentiated from RANKL-stimulated RAW264.7 cells, induced pit formation with a longer time of EPR in a gefitinib (an EGFR tyrosinekinase inhibitor)-sensitive manner. These findings suggest that both EPR and EGF acted on osteoblasts to enhance bone resorption indirectly. In addition, EPR is suggested to have a direct stimulating pathway through the activation of EGFR on mature osteoclasts leading to bone resorption.

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Nuclear factor E2-related factor 2 (nrf2) is accepted as an important transcription factor in the regulation of many phase II detoxifying enzymes and oxidative-stress-inducible genes in many tissues including bone and cartilage. However, little attention has been paid to precise role of nrf2 on osteoblasts and chondrocytes. In the present study, we therefore investigated the role of nrf2 in the regulation of cell proliferation, differentiation and maturation using both MC3T3-E1 and ATDC5 cells. In MC3T3-E1 cells stably transfected with nrf2 (MC3T3-E1-nrf2 clone), the differentiation-dependent induction of alkaline phosphatase activity was significantly inhibited in addition to decrease of the mineralized matrix formation. Moreover, expression of mRNAs for osteocalcin and type I collagen was also markedly attenuated in MC3T3E1-nrf2 clone but not in MC3T3E1-empty vector clone. These inhibitory effect on cell differentiation was also observed in ATDC5 cells stably transfected with nrf2. Moreover, nrf2 significantly impaired the runx2-dependent enhancement of osteocalcin promoter activity. These data suggest that nrf2 may negatively regulate the differentiation of both osteoblasts and chondrocytes through inhibition of the runx2 dependent transcriptional activity.

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A new injectable biomimetic composite cement composed of nano-hydroxyapatite (n-HA) and polyamide 66 (polyhexamethylene adipamide) has been developed. This study investigated that in vitro biomechanical performances of three n-HA/PA composite cements in the augmentation of osteoporotic cancellous bone were different in various n-HA content to evaluate the clinical applicability. The thoracic vertebrae and femoral condyles of five osteoporotic and one normal cadaver were utilized to divide into five groups: 60wt%, 70wt%, 80wt%, the osteoporotic and the normal groups. The thoracic vertebrae performed by vertebroplasty were loaded in compression until 75 percent of vertebral height. The cancellous bone specimens made of femoral condyles were applied to torsional testing. Biomechanical testing showed the compressive and torsional performances in the groups injected with 60%, 70% and 80% n -HA/PA composite cement increased markedly than that in osteoporosis. Some of the mechanical parameters were close to normal cancellous bone, in particular, 70% group in compressive properties and 60% group in torsional properties. This study demonstrated that the injectable n-HA/PA composite cement was capable of strengthening the compressive and torsional properties of osteoporotic cancellous bone and suggested that 60% and 70% cement might be competent materials in the treatment and prophylaxis of osteoporotic fracture.

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Vitamin K (K) is essential for blood coagulation and bone metabolism in mammals. K acts as a cofactor in the posttranslational synthesis of g-carboxyglutamic acid from glutamic acid residues. In addition to liver and bone, K is found in brain, heart, kidney and gonadal tissue. However, the physiological role of K in these varied organs is not yet fully understood. It is likely that K has functions in addition to its role as a cofactor of protein g-glutamyl carboxylation. In this paper we used DNA microarray techniques to identify the effect of K status on gene expression in rat liver. Expression of genes involved in the acute inflammation response was enhanced in rats fed a K-deficient diet relative to control and K₁-supplemented diet groups. Moreover, dietary supplementation with K₁ suppressed inflammation induced by LPS administration. These results indicate that orally administrated K₁ suppresses inflammation in the rat.

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The objective of this study is to develop an osteoinductive scaffold which enables the controlled release of bone morphogenetic protein (BMP)-2. A biodegradable gelatin sponge incorporating 50 wt% of b-tricalcium phosphate (β-TCP) was fabricated to investigate the osteoinduction activity. The sponge prepared had an interconnected pore structure with an average pore size of 200 mm. The in vivo release test revealed that BMP-2 was released from the sponge for a time period longer than 28 days. Combination of the sponge capable for BMP-2 release and autologous bone marrow cells significantly induced bone regeneration at a segmental defect of rabbit ulnae. It is concluded that the gelatin sponge for BMP-2 release is a promising cell scaffold for osteoinduction in vivo.

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Recently, platelet-rich plasma (PRP) has been clinically employed to promote bone repairing. However, little has been investigated on the materials combination of PRP to enhance the biological function of growth factors present in platelets. In this study, feasibility of gelatin hydrogels in the controlled release of platelet growth factors and the consequent enhancement of PRP-induced bone regeneration were evaluated by a cranial bone defect model of rabbits. In conclusion, the gelatin hydrogel achieved the controlled release of bioactive platelet growth factors to significantly promote bone regeneration, in marked contrast to PRP alone.

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Human papilloma viruses (HPV) infection is a significant risk factor for uterine cervical carcinoma. Many previous studies have also demonstrated the presence of HPVs in oral epithelia tissue. However, the role of HPV infection in oral squamous cell carcinoma (OSCC) is still controversy. The present study is to determine the frequency and type of HPV in OSCCs and oral pre-cancer lesions. Methods: DNA samples were collected from 51 OSCCs, 46 oral pre-cancer lesions and 90 normal control specimens by cytobrushings. Nested polymerase chain reaction (PCR), and gene-chip are used to identify multiple HPV types in our samples. Results: The positive rates of overall HPV types (14/46, P=0.0216, OR=2.844, CI=1.186-6.816) and of low-risk types (9/46, P=0.0096, OR=5.529, CI=1.597-19.14) are significantly higher in oral pre-cancer lesions than in control samples. The prevalence of high-risk type (11/51, P=0.0420, OR=2.819, CI=1.051-7.558) is significantly higher in OSCCs than in control but of overall types (13/51, P=0.1066, OR=2.244, CI=0.9266-5.337) is not to reach the statistical significance. Conclusion: The high-risk HPV may play a role in OSCCs progression and the low-risk ones may associate with the oral pre-cancer lesions.

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Preliminary analysis of cells before using them in tissue engineering is mandatory. Thus, to clarify the behavior of cells in an appropriate experimental model, we evaluated the characteristic of MDPC-23 cells and KUSA/A1 cells in vitro and in vivo seeded in diffusion chamber. Our results indicated that KUSA/A1 cells differentiated into osteoblast-like cells and induced bone tissue inside the chamber. Whereas, MDPC-23 cells were odontoblast-like cells but with low ability to induce dentin formation. This study suggests that MDPC-23 cells are special cells, which possess morphological and functional character of odontoblast-like cells, expressing DSPP only in vivo with low capacity to induce mineralized dentin matrix.

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We previously reported the characterization of the genomic structure of the human ING1 gene and found tumor specific missense mutations. We also demonstrated that four mRNA variants were transcribed from three different promoter regions. In this study, we examined the mRNA expression of two major splicing variants of ING1 gene in 40 matched samples of normal and tumor tissues from head and neck cancers by RT-PCR. One of the splicing variant, p24, showed decreased and increased expression in 51% and 27% of the samples, respectively, while another major variant, p33, demonstrated low and high mRNA expression in 20% and 35% of the samples, as compared to normal controls. To elucidate the silencing mechanism of the ING1 gene, we examined the methylation status of each splicing variant. Our results suggest that alternative variants of ING1 gene may have different role in the carcinogenic pathway of head and neck cancers.

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Loss of heterozygosity (LOH) has been frequently detected at chromosome 7q31 region in human head and neck squamous cell carcinomas (HNSCC) and many other cancers, suggesting the existence of the tumor suppressor genes (TSG). However, the targeted gene was yet identified. We analyzed LOH at 7q31 region in 49 HNSCC by using 6 polymorphic microsatellite markers and found allelic deletion in 48% of the informative cases. We detected two preferentially deleted locations one around D7S643 and the other around D7S486. When we redefined the map of 7q31 region according to the contiguous sequences, an ING1 like gene, ING3, which we named it due to homology to ING1, was found in the proximity of D7S643. ING3 protein harbors a PHD zinc finger domain highly homologous among ING family proteins, in which we previously found mutations in a related gene, ING1. As only one missense mutation of the ING3 gene was found in HNSCC, we examined the mRNA expression levels. RT-PCR analysis demonstrated decreased or no expression of ING3 mRNA in 45% of primary tumors as compared with that of mathced normal samples. All these findings suggest a possibility that ING3 functions as a TSG in a subset of HNSCC.

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We previously defined human chromosome 10q21 as a hotspot of regional loss in head and neck squamous cell carcinomas (HNSCC) by genome-wide loss of heterozygosity (LOH) analysis. Aims of this study are to narrow-down the target area by using new microsatellite markers and to define candidate tumor suppressor genes (TSG). LOH analysis on 10q21 in 52 HNSCC by 8 highly polymorphic markers indicated distinctive and frequent allelic loss at D10S589 (42%). Among flanking genes, we found the RHOBTB1 gene as a candidate TSG, since an intragenic marker demonstrated the highest LOH (44%). Semi-quantitative expression analysis revealed down-regulation of RHOBTB1 mRNA in 37% of tumors. Interestingly, all tumors that showed decreased expression of RHOBTB1 were accompanied with LOH, supporting the haploinsufficiency and class 2 TSG characteristics of RHOBTB1. Although no pathogenic mutation of RHOBTB1 was found, frequent allelic loss and decreased expression of RHOBTB1 suggested that this gene has a role in tumorigenesis of a subset of HNSCC.

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Biochemical and genetic studies has identified several ATP-dependent multiprotein complexes that are involved in the remodeling of chromatin during gene activation. The SWI/SNF complex is one of these molecules. Molecular pathological changes of several members of SWI/SNF complex, especially human brahma (hBRM) and brahma-related gene 1 (BRG1) were reported to be involved in carcinogenesis of human cancer. hBRM can bind and cooperate with hypophosphorylated RB protein in repressing E2F1 transcriptional activation in transient transfection studies. We analyzed the LOH of the short arm of chromosome 9 in 64 head and neck squamous cell carcinomas by using 13 highly polymorphic microsatellite markers and found two deletion hot spots at 9p21 and 9p24. P16 tumor suppressor gene is likely to be a target for the deletion of 9p21 region. When the map of 9p24 region was redefined, a possible tumor suppressor gene hBRM was identified. Therefore we prepared an hBRM specific microsatellite marker and found 67% deletion of this gene at 9p24 region. In RT-PCR analysis about 60% of tumor samples demonstrated reduced mRNA expression as compared to matched normal samples.

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Oral Squamous Cell Cancer is the most common malignant tumour in head and neck area being the top of Oral Carcinoma. The malignancy is high, invasion is strong, and metastasis is fast. This report will use the immunohistochemical method to test the expression of MMP-2 and TIMP-2 in normal oral mucosa, dysplastic oral mucosa and oral squamous tissue, in order to investigate the correlativity and the effect to the biological behaviour of Oral Squamous Cell Cancer. In the process of invasion and metastasis of carcinoma, imbalance of MMP and TIMP expression plays an important role. Detecting MMP and TIMP the protein expression may be of great significance to estimating the biological behavior of carcinoma and evaluating the prognosis of patients.

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By the tsunami occurred in the Indian Ocean at the end of last year, more than 200,000 of human life were lost. Since there are many famous resorts in the stricken area, a lot of overseas' tourists were also killed by that tsunami.
It is well known that dentists were dispatched from many countries and they performed victim's identifications by the method of the dental findings. Japan also sent several dentists to Thailand and Indonesia respectively. In Japan, more than 90 percent of people have a record of dentistry consultation, therefore, the dental findings are able to work as a very effective means for personal identification. Under such circumstances, there are six laboratories concerned to forensic odontology among 28 dental schools and in the most of those schools; forensic odontology is lectured to the students.
Furthermore, dental associations in cooperation with police are organized in every prefectures in Japan and it is established such system as they promptly set out and cooperate with police, once disaster or criminal case should be occurred. Dentist as a member of the said associations regularly receive the training how to take the dental findings and confirm the identity of person at postmortem inspection because it is thought that the skill of dentist to do the personal identification can be improved by the practice of daily cases.

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The short tandem repeat (STR) polymorphisms are highly useful tools for personal identification and paternity testing in forensic practice. Sequence analysis of mitochondrial DNA (Mt DNA) is being used to characterize forensic biological specimens, particularly when there is insufficient nuclear DNA in samples for typing. Bones, teeth and other samples that are severely decomposed may be subjected to Mt DNA analysis. We could successfully apply these techniques to investigate samples from practical forensic cases and confirming their efficacy for forensic practice.

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To develop submandibular gland acellular matrix and observe its structure and analyze its histological ingredients. METHODS: The fresh submandibular glands of SD mice were cell-extracted by chemical detergent and histological ingredients were analyzed by immunohistochemistry and ultra structure observed. RESULTS: Submandibular gland cells disappeared, matrix ingredients appeared network structure under microscopy and The collagen fibers showed different diameter and directions and formed complicated network structure and SEM. I, II and III type collagen proteins of submandibular gland before and after cell-extracted were positive under immunohistochemistry. CONCLUSIONS: The main ingredients of submandibular gland acellular matrix were I, II and III type collagen proteins.

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A candidate tumor suppressor gene, ING1 has been recently cloned and mapped to 13q33-34 region. We characterized the genomic structure of the human ING1 gene and detected somatic mutations of the ING1 gene in head and neck squamous cell carcinomas. 23 out of 34 informative cases (68%) of tumors showed loss of heterozygosity at chromosome 13q33-34, where the ING1 gene is located. By sequence analysis three exons and 2 introns of ING1 were identified. Mutation analysis of ING1 gene showed three missense and three silent changes.

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We previously showed two members of the ING family, ING1 and ING3 as a tumor suppressor gene in head and neck cancer. A novel member of ING family, ING4 localized to chromosome 12p13.31 region was cloned. ING4 harbors the PHD domain highly homologous among ING family proteins. Loss of heterozygosity analysis was performed at 12p12-13 region in 50 head and neck squamous cell carcinomas by using six highly polymorphic microsatellite markers and 33 out of the 50 cases (66%) showed allelic loss. Although somatic mutation of the ING4 gene was not found in head and neck cancers, quantitative real-time RT-PCR analysis demonstrated decreased expression of ING4 mRNA in 76% of primary tumors as compared with that of matched normal samples. Frequent deletion and decreased mRNA expression of ING4 suggested it as a class II tumor suppressor gene and may play an important role in head and neck cancer.

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We analyzed loss of heterozygosity in detail of the long arm of chromosome 4 in 40 oral cancer by using 16 microsatellite markers based on the recent data from human genome sequence and defined the deletional mapping of the region with putative tumor suppressor genes. Our data revealed two distinct commonly deleted regions around the markers D4S2623 and D4S1644 with an allelic deletion of 44% and 39%, respectively. Additional mapping and use of the markers near one of these hot spots narrowed down the minimally deleted region about 1.5 Mbp around the marker, D4S2623. Caspase 6 is just localized 280 kb distant from the marker, D4S2623. Fine mapping of this region with possible tumor suppressor gene suggest caspase 6 as a putative tumor suppressor gene. Further molecular analysis of caspase 6 should be performed to clarify its role in oral carcinogenesis.

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We recently reported the frequent deletion of chromosome 4q25 region in oral squamous cell carcinoma and discussed the Caspase-6 as a possible tumor suppressor gene in this region due to its near location to this hot spot. We this time examined the role of Caspase-6 in oral cancers. mRNA expression level, protein localization and gene mutation of Caspase-6 were analyzed. Reverse-transcription-PCR analysis demonstrated decreased expression in 40% and equal expression in 30% of Caspase-6 mRNA of primary tumors as compared with that matched normal samples. cDNA sequencing analysis of Caspase-6 did not reveal any somatic mutation. Caspase-6 protein localization was in similar proportion with its mRNA expression. Failure of Caspase-6 response in tumor samples as compared to normal samples could suggest Caspase-6 as a candidate tumor suppressor gene in a subset of oral cancer.

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To detect deletions of specific chromosome regions, loss of heterozygosity (LOH) analysis is a sensitive method. In different types of human cancer, BRG1, a member of SWI/SNF complex proteins, located at 19p13.2, suggested to be a candidate tumor suppressor gene (TSG). We detected allelic deletion in 25 of 39 (64%) samples at 19p13 by using six microsatellite markers. The BRG1 specific microsatellite marker showed the highest LOH in tumor samples. As we couldn't detect any mutation of the BRG1 gene in oral cancers, we examined the mRNA expression level. During expression analysis we detected an alternative in frame splicing form of BRG1, which includes exon 26 is selectively decreased or lost in most tumor samples. This 33 aminoacid sequence of BRG1 protein shows very high homology with heterogenous nuclear ribonucleoprotein E. Thus may affect the function and level of BRG1 through modifications on post-transcriptional control.

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We demonstrate that a new culture system using a titanium web (TW) consisted of titanium fibers (50 mm diameter) can promote differentiation of mesenchymal cells into osteoblasts without conventional application of dexamethasone. Human mesenchymal stem cells (hMSC, Cambrex) and MC3T3-E1 were cultured on titanium web (87% of porosity). ALP/DNA in hMSC at 2 weeks was 2 times higher and MC3T3-E1 at 4 weeks was 4 times higher on TW than those on the conventional plastic dishes. Effect of 3D structure of TW is self-evident and the results will open a new age of 3D bone cell biology using TW.

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