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Construction and characterization of Human oral mucosal equivalent using Amniotic membrane as a matrix
Poster Session in the co-cultured C3H10T1/2 cells than in monocultured C3H10T1/2 cells. Conclusions: The present study implies that osteoblasts and MSCs communicate via gap junctions. Furthermore, our results strongly suggest that chromatin remodeling followed by ALP and BSP transcription occur in MSCs as a result of gap junction-mediated intercellular communication. This indicates that osteoblasts induce osteogenic differentiation of MSCs. To the best of our knowledge, this is the first demonstration of gap junction-modulated transcription in stem cells. While these observations have enormous implications for understanding the mechanisms of bone remodeling and MSCs maintenance and differentiation. References: 1. Mikami Y, Matsumoto T, Kano K, Toriumi T, Somei M, Honda MJ, Komiyama K. (2014) Current status of drug therapies for osteoporosis and the search for stem cells adapted for bone regenerative medicine. Anat Sci Int 89:1-10. 2. Marotti G and Palumbo C. (2001) The mechanism of transduction of mechanical strains into biological signals at the bone cellular level. Eur J Histochem 51:15-19.
POSTER 287 Virtual Surgical Planning in the Era of Robotic Surgery J. H. Park: College of Dentistry, Yonsei University, J. Y. Kim, W. Nam, J. G. Kim Robot assisted neck dissection offers more esthetic outcomes. Virtual surgical planning (VSP), a method of pre-visualizing a surgical intervention with computer simulation, can help robotic surgery with saved time and allow esthtic reconstructions with customized osteotomy lines. Objective: To discuss about efficacy of virtual surgical planning (VSP) in the era of robotic surgery. Methods: Two mandibular reconstruction cases after robotic neck dissection using the conventional method and virtual surgical planning (VSP), respectively. Results: We found the conventional method limits surgical scope, restricts access to the defect area, and consumes considerable time. Conclusions: The authors consider VSP far more viable in the era of robotic surgery. References: 1. Yang X, Hu J, Zhu S, Liang X, Li J, Luo E. Computer-assisted surgical planning and simulation for condylar reconstruction in patients with osteochondroma. Br J Oral Maxillofac Surg 2011;49:203-8. 2. Zheng GS, Su YX, Liao GQ, Chen ZF, Wang L, Jiao PF, et al. Mandible reconstruction assisted by preoperative virtual surgical simulation. Oral Surg Oral Med Oral Pathol Oral Radiol 2012;113: 604-11.
AAOMS 2014
POSTER 288 Construction and characterization of Human oral mucosal equivalent using Amniotic membrane as a matrix F. Qi: Shinshu University School of Medicine, T. Shimane, H. Aizawa, Y. Li, H. Kurita Statement of the Problem: A clinical need exists for an immunologically compatible surgical patch for the use of soft tissue replacement, body wall repair, and as a wound dressing. Amniotic membrane is a thin lightweight and elastic material and it can promote cell attachment, proliferation and differentiation1. Hyper-dry amniotic membrane as a natural extracellular matrix is a novel dried amniotic, can be preserved at room temperature2and has been used to culture epithelial cell for transplantation. In this study, we made human oral mucosal equivalent using hyper-dry amniotic membrane as a matrix. The cultured oral mucosal equivalent was transplanted in mouse full-thickness skin defect, the wound contraction and characterization of the grafted material was evaluated. Materials and Methods: Gingival mucosa was obtained from wisdom teeth extraction patient. Keratinocytes extracted from gingival mucosa and developed in a serum-free culture system without a feeder layer. Oral keratinocytes were seeded and cultured on hyper-dry amniotic membrane (HAM, Toyama University) for two weeks. Full-thickness skin wounds (1 cm x 1 cm) were made on athymic mice (BALB/C) and covered with the oral mucosal equivalent cultured on hyper-dry AM (OME on HAM), with the oral mucosal equivalent cultured on artificial membrane (MillcellR culture plate insert) , and with no material offers as control. 3 weeks after grafting, constriction of wound and characterization of grafted material were examined with hematoxylin and eosin and immunohistochemcial staining for ck10, ck1, and involucrin. Results: By two weeks that Keratinocytes cultured on HAM revealed 4 to 5-layered well-organized stratified epithelium on the surface. Presence of ck10, ck1 in all suprabasal layers , While involucrin was presence in all cell layers. By three weeks of mice grafting with OME on HAM, the regenerated oral mucosal showing a good morphology which is similar to gingival mucosa and the epithelial cells expressed ck10, ck1 and involucrin in supra-basal layers is similar to that of gingival mucosa. Concerning wound contraction, OME on HAM showed a smallest contraction rate (87.97 2.39 % with OME on HAM, 89.111.66%with OME on MillcellR, 91.232.1% in control). Conclusions: Hyperdry AM have sufficient elasticity and can be cut in a suitable shape easily. This study demonstrated that use Hyper-dry AM as a matrix developed oral
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Poster Session mucosal equivalent revealed a good morphology and contribute to a stable wound healing. References: 1. Lujun Yang, Yuji Shirakata, Sho Tokumara. Living skin equivalents constructed using human amnions as a matrix. Journal of Dermatological Science 56, (2009) 188-195 2. Ayaka Toda, Motonori Okabe, Toshiko Yoshida. The potential of amniotic membrane/amnion-derived cells for regeneration of Various Tissues. Journal of Pharmacological sciences. 105, (2007) 215-228
POSTER 289 A Comparative Study of The Effectiveness of Platelet Rich Fibrin on Bone Regeneration of Various Grafts in Sinus Floor Augmentation E. Peker: Gazi University Faculty of Dentistry, I. Karaca, Y. Yildiz, B. Yildirim The objective of this study was to evaluate the effectiveness of various grafting materials used for augmentation of the maxillary sinus floor in a rabbit model. 36 adult New Zealand white rabbits were used in this study. Bilateral augmentation of the floor of the maxillary sinus was performed with autogenous bone or other materials. The rabbit maxillary sinuses divided into 4 groups, one of the following was grafted to the maxillary sinus of each rabbit: Demineralized bone matrix (DBM) [Grafton (Osteotech, Eatontown, NJ)] DBM combined with platelet-rich-fibrin (PRF), collagenated heterologous bone graft [Apatos Mix (OsteoBiolÒ, TecnossÒ, Coazze, Italy)] and collagenated heterologous bone graft combined with PRF. Animals were sacrificed at 2nd, 4th and 8thweeks after surgery. Histological and immunohistochemical examinations were performed to assess biological healing process in the various samples. Hematoxylin-eosin staining was performed for evaluation of bone formation and bone morphogenic protein (BMP) antibody staining was performed for the immunohistochemical evaluation. The data obtained from this study were evaluated with SPSS 20.0 software package. For statistical analysis, Mann Whitney U test and Kruskal Wallis H test with Bonferroni correction were used to examine the differences between the groups. In all groups new bone formation was seen at 2 to 8 weeks. Statistical analysis of new bone formation revealed no significant differences between PRF+DBM group and PRF+collagenated heterologous bone graft.(p>0.05) According to statistical analysis, total area of residual graft particles and percentage of this area is higher in collagenated heterologous bone graft group than DBM group.(p<0.05) Total new bone area and percentage of this area is higher in the DBM group than e-214
collagenated heterologous bone graft group.(p<0.05) According to the immunohistochemical examinations; BMP staining cells are slightly increased in all groups from 2 to 8 weeks, but there is no significant difference in staining of BMP antibody between all groups in the 2nd, 4th and 8thweeks.(p>0.05) This study showed no positive effect of PRF on new bone formation and BMP expression when combined with demineralized bone matrix and collagenated heterologous bone graft according to the histological and immonuhistochemical evaluation when used in sinus floor augmentation. Demineralized bone matrix improves the outcomes of the sinus augmentation when compared to collagenated heterologous bone graft. References: 1. Choukroun J, Diss A, Simonpieri A, Girard MO, Schoeffler C, Dohan SL, et al. Platelet-rich fibrin (PRF): A second-generation platelet concentrate. Part V: Histologic evaluations of PRF effects on bone allograft maturation in sinus lift. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2006;101:299–303. 2. Yu Zhang, Stefan Tangl, Christian D Huber, Ye Lin, Lixin Qiu, Xiaohui Rausch-Fan. Effects of Choukroun’s platelet-rich-fibrin on bone regeneration in combination with deproteinized bovine bone mineral in maxillary sinus augmentation: a histological and histomorphometric study. J Craniomaxillofac Surg. 2012 Jun;40(4):321-8.
POSTER 290 Donor site morbidity following Mono cortical Deep Circumflex Iliac Artery flap S. Y. Moon: School of Dentistry, Chosun University Statement of the Problem: Conventionally deep circumflex iliac artery (DCIA) flap had been harvested as bicortical form. However, the result of this procedure, several complications and side effects occurred such as abnormal hip contour, hernia, severe bleeding tendency, gait disturbance and hypoesthesia. The aim of this study is to evaluate donor site morbidity of monocortical DCIA flap for reconstruction of jaw defect. Subjects and Methods: All of 7 patients who required reconstruction of the jaw with microvascular free flaps after radical resection. Patients were assessed donor site morbidity by asking about pain and sensory disturbance, and availability of normal activities. Results: Monocortical bone was harvested from the anterior iliac crest and the amount of bone harvested was from 47mm to 90mm (mean 6314.6). The number of days to feel subjective spontaneous pain after surgery were from 10 to 30 days, however patient could walk after from 5 to 14 days (8.33.1). Resumed normal daily activities, including running within 60 days. Conclusions: The present study has confirmed that low donor site morbidity relating to the gait disturbance is associated with reconstruction with monocortical DCIA flap. All the patients were able to resume normal daily activities within 2 months. AAOMS 2014
POSTER 287 Virtual Surgical Planning in the Era of Robotic Surgery J. H. Park: College of Dentistry, Yonsei University, J. Y. Kim, W. Nam, J. G. Kim Robot assisted neck dissection offers more esthetic outcomes. Virtual surgical planning (VSP), a method of pre-visualizing a surgical intervention with computer simulation, can help robotic surgery with saved time and allow esthtic reconstructions with customized osteotomy lines. Objective: To discuss about efficacy of virtual surgical planning (VSP) in the era of robotic surgery. Methods: Two mandibular reconstruction cases after robotic neck dissection using the conventional method and virtual surgical planning (VSP), respectively. Results: We found the conventional method limits surgical scope, restricts access to the defect area, and consumes considerable time. Conclusions: The authors consider VSP far more viable in the era of robotic surgery. References: 1. Yang X, Hu J, Zhu S, Liang X, Li J, Luo E. Computer-assisted surgical planning and simulation for condylar reconstruction in patients with osteochondroma. Br J Oral Maxillofac Surg 2011;49:203-8. 2. Zheng GS, Su YX, Liao GQ, Chen ZF, Wang L, Jiao PF, et al. Mandible reconstruction assisted by preoperative virtual surgical simulation. Oral Surg Oral Med Oral Pathol Oral Radiol 2012;113: 604-11.
AAOMS 2014
POSTER 288 Construction and characterization of Human oral mucosal equivalent using Amniotic membrane as a matrix F. Qi: Shinshu University School of Medicine, T. Shimane, H. Aizawa, Y. Li, H. Kurita Statement of the Problem: A clinical need exists for an immunologically compatible surgical patch for the use of soft tissue replacement, body wall repair, and as a wound dressing. Amniotic membrane is a thin lightweight and elastic material and it can promote cell attachment, proliferation and differentiation1. Hyper-dry amniotic membrane as a natural extracellular matrix is a novel dried amniotic, can be preserved at room temperature2and has been used to culture epithelial cell for transplantation. In this study, we made human oral mucosal equivalent using hyper-dry amniotic membrane as a matrix. The cultured oral mucosal equivalent was transplanted in mouse full-thickness skin defect, the wound contraction and characterization of the grafted material was evaluated. Materials and Methods: Gingival mucosa was obtained from wisdom teeth extraction patient. Keratinocytes extracted from gingival mucosa and developed in a serum-free culture system without a feeder layer. Oral keratinocytes were seeded and cultured on hyper-dry amniotic membrane (HAM, Toyama University) for two weeks. Full-thickness skin wounds (1 cm x 1 cm) were made on athymic mice (BALB/C) and covered with the oral mucosal equivalent cultured on hyper-dry AM (OME on HAM), with the oral mucosal equivalent cultured on artificial membrane (MillcellR culture plate insert) , and with no material offers as control. 3 weeks after grafting, constriction of wound and characterization of grafted material were examined with hematoxylin and eosin and immunohistochemcial staining for ck10, ck1, and involucrin. Results: By two weeks that Keratinocytes cultured on HAM revealed 4 to 5-layered well-organized stratified epithelium on the surface. Presence of ck10, ck1 in all suprabasal layers , While involucrin was presence in all cell layers. By three weeks of mice grafting with OME on HAM, the regenerated oral mucosal showing a good morphology which is similar to gingival mucosa and the epithelial cells expressed ck10, ck1 and involucrin in supra-basal layers is similar to that of gingival mucosa. Concerning wound contraction, OME on HAM showed a smallest contraction rate (87.97 2.39 % with OME on HAM, 89.111.66%with OME on MillcellR, 91.232.1% in control). Conclusions: Hyperdry AM have sufficient elasticity and can be cut in a suitable shape easily. This study demonstrated that use Hyper-dry AM as a matrix developed oral
e-213
Poster Session mucosal equivalent revealed a good morphology and contribute to a stable wound healing. References: 1. Lujun Yang, Yuji Shirakata, Sho Tokumara. Living skin equivalents constructed using human amnions as a matrix. Journal of Dermatological Science 56, (2009) 188-195 2. Ayaka Toda, Motonori Okabe, Toshiko Yoshida. The potential of amniotic membrane/amnion-derived cells for regeneration of Various Tissues. Journal of Pharmacological sciences. 105, (2007) 215-228
POSTER 289 A Comparative Study of The Effectiveness of Platelet Rich Fibrin on Bone Regeneration of Various Grafts in Sinus Floor Augmentation E. Peker: Gazi University Faculty of Dentistry, I. Karaca, Y. Yildiz, B. Yildirim The objective of this study was to evaluate the effectiveness of various grafting materials used for augmentation of the maxillary sinus floor in a rabbit model. 36 adult New Zealand white rabbits were used in this study. Bilateral augmentation of the floor of the maxillary sinus was performed with autogenous bone or other materials. The rabbit maxillary sinuses divided into 4 groups, one of the following was grafted to the maxillary sinus of each rabbit: Demineralized bone matrix (DBM) [Grafton (Osteotech, Eatontown, NJ)] DBM combined with platelet-rich-fibrin (PRF), collagenated heterologous bone graft [Apatos Mix (OsteoBiolÒ, TecnossÒ, Coazze, Italy)] and collagenated heterologous bone graft combined with PRF. Animals were sacrificed at 2nd, 4th and 8thweeks after surgery. Histological and immunohistochemical examinations were performed to assess biological healing process in the various samples. Hematoxylin-eosin staining was performed for evaluation of bone formation and bone morphogenic protein (BMP) antibody staining was performed for the immunohistochemical evaluation. The data obtained from this study were evaluated with SPSS 20.0 software package. For statistical analysis, Mann Whitney U test and Kruskal Wallis H test with Bonferroni correction were used to examine the differences between the groups. In all groups new bone formation was seen at 2 to 8 weeks. Statistical analysis of new bone formation revealed no significant differences between PRF+DBM group and PRF+collagenated heterologous bone graft.(p>0.05) According to statistical analysis, total area of residual graft particles and percentage of this area is higher in collagenated heterologous bone graft group than DBM group.(p<0.05) Total new bone area and percentage of this area is higher in the DBM group than e-214
collagenated heterologous bone graft group.(p<0.05) According to the immunohistochemical examinations; BMP staining cells are slightly increased in all groups from 2 to 8 weeks, but there is no significant difference in staining of BMP antibody between all groups in the 2nd, 4th and 8thweeks.(p>0.05) This study showed no positive effect of PRF on new bone formation and BMP expression when combined with demineralized bone matrix and collagenated heterologous bone graft according to the histological and immonuhistochemical evaluation when used in sinus floor augmentation. Demineralized bone matrix improves the outcomes of the sinus augmentation when compared to collagenated heterologous bone graft. References: 1. Choukroun J, Diss A, Simonpieri A, Girard MO, Schoeffler C, Dohan SL, et al. Platelet-rich fibrin (PRF): A second-generation platelet concentrate. Part V: Histologic evaluations of PRF effects on bone allograft maturation in sinus lift. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2006;101:299–303. 2. Yu Zhang, Stefan Tangl, Christian D Huber, Ye Lin, Lixin Qiu, Xiaohui Rausch-Fan. Effects of Choukroun’s platelet-rich-fibrin on bone regeneration in combination with deproteinized bovine bone mineral in maxillary sinus augmentation: a histological and histomorphometric study. J Craniomaxillofac Surg. 2012 Jun;40(4):321-8.
POSTER 290 Donor site morbidity following Mono cortical Deep Circumflex Iliac Artery flap S. Y. Moon: School of Dentistry, Chosun University Statement of the Problem: Conventionally deep circumflex iliac artery (DCIA) flap had been harvested as bicortical form. However, the result of this procedure, several complications and side effects occurred such as abnormal hip contour, hernia, severe bleeding tendency, gait disturbance and hypoesthesia. The aim of this study is to evaluate donor site morbidity of monocortical DCIA flap for reconstruction of jaw defect. Subjects and Methods: All of 7 patients who required reconstruction of the jaw with microvascular free flaps after radical resection. Patients were assessed donor site morbidity by asking about pain and sensory disturbance, and availability of normal activities. Results: Monocortical bone was harvested from the anterior iliac crest and the amount of bone harvested was from 47mm to 90mm (mean 6314.6). The number of days to feel subjective spontaneous pain after surgery were from 10 to 30 days, however patient could walk after from 5 to 14 days (8.33.1). Resumed normal daily activities, including running within 60 days. Conclusions: The present study has confirmed that low donor site morbidity relating to the gait disturbance is associated with reconstruction with monocortical DCIA flap. All the patients were able to resume normal daily activities within 2 months. AAOMS 2014