Khang, Gilson;Rhee, John M.;Shin, Philkyung;Kim, In Young;Lee, Bong;Lee, Sang Jin;Lee, Young Moo;Lee, Hai Bang;Lee, Ilwoo
Macromolecular Research
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제10권3호
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pp.158-167
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2002
In order to endow with new bioactive functionality from small intestine submucosa (SIS) powder as natural source to poly (L-lactide) (PLA) and poly (lactide-co-glycolide) (PLGA) synthetic biodegradable polymer, porous SIS/PLA and SIS/PLGA as natural/synthetic composite scaffolds were prepared by means of the solvent casting/salt leaching methods for the possibility of the application of tissue engineered bone and cartilage. A uniform distribution of good interconnected pores from the surface to core region was observed the pore size of 40~500 ${\mu}{\textrm}{m}$ independent with SIS amount using the solvent casting/salt leaching method. Porosities, specific pore areas as well as pore size distribution also were almost same. After the fabrication of SIS/PLA hybrid scaffolds, the wetting properties was greatly enhanced resulting in more uniform cell seeding and distribution. Five groups as PGA non-woven mesh without glutaraldehyde (GA) treatment, PLA scaffold without or with GA treatment, and SIS/PLA (Code No.3 ; 1 : 12 of salt content, (0.4 : 1 of SIS content, and 144 ${\mu}{\textrm}{m}$ of median pore size) without or with GA treatment were implanted into the back of nude mouse to observe the effect of SIS on the induction of cells proliferation by hematoxylin and eosin, and von Kossa staining for 8 weeks. It was observed that the effect of SIS/PLA scaffolds with GA treatment on bone induction are stronger than PLA scaffolds, that is to say, in the order of PLA/SIS scaffolds with GA treatment > PLA/SIS scaffolds without GA treatment > PGA nonwoven > PLA scaffolds only with GA treatment = PLA scaffolds only without GA treatment for the osteoinduction activity. The possible explanations are (1) many kinds of secreted, circulating, and extracellular matrix-bound growth factors from SIS to significantly affect critical processes of tissue development and differentiation, (2) the exposure of SIS to GA resulted in significantly calcification, and (3) peri-implant fibrosis due to covalent bonding between collagen molecule by crosslinking reaction. In conclusion, it seems that SIS plays an important role for bone induction in SIS/PLA scaffolds for the application of tissue engineering area.
Although the main purpose of periodontal treatment to regenerate is the complete regeneration of periodontal tissue due to periodontal disease, most of the treatment cannot meet such purpose because healing by long epithelial junction. Therefore, diverse materials of resorbable and non-resorbable have been used to regenerate the periodontal tissue. Due to high risk of exposure and necessity of secondary surgical procedure when using non-resorbable membrane, guided tissue regeneration using the resorbable membrane has gain popularity, recently. However, present resorbable membrane has the disadvantage of not having sufficient time to regenerate date to the difference of resorption rate according to surgical site. Meanwhile, other than the structure stability and facile manipulation, acellular dermal matrix has been reported to be a possible scaffold for cellular proliferation due to rapid revascularization and favorable physical properties for cellular attachment and proliferation. The purpose of this study is to estimate the influence of acellular dermal matrix on periodontal ligament, cementum and alveolar bone when acellular dermal matrix is implanted to 1-wall alveolar bone defect. 4 dogs of 12 to 16 month old irrelevant to sex , which below 15Kg of body weight, has been used in this study. ADM has been used for the material of guided tissue regeneration. The 3rd premolar of the lower jaw was extracted bilaterally and awaited for self-healing. subsequently buccal and lingual flap was elevated to form one wall intrabony defect with the depth and width of 4mm on the distal surface of 2nd premolar and the mesial surface of 4th premolar. After the removal of periodontal ligament by root planing. notch was formed on the basal position. Following the root surface treatment, while the control group had the flap sutured without any treatment on surgically induced intrabony defect. Following the root surface treatment, the flap of intrabony defect was sutured with the ADM inserted while the control group sutured without any insertion. The histologic specimen was observed after 4 and 8 weeks of treatment. The control group was partially regenerated by periodontal ligament, new cementum and new alveolar bone. the level of regeneration is not reached on the previous formed notch. but, experimental group was fully regenerated by functionally oriented periodontal ligament fiber. new cementum and new alveolar bone. In conclusion, we think that ADM seems to be used by scaffold for periodontal ligament cells and the matrix is expected to use on guided tissue regeneration.
Epithelial-mesenchymal interaction plays a important role in cell growth and differentiation. This interaction is already well known to have an importance during the organ development as well as cell growth and differentiation. However, in vitro experimental model is not well developed to reproduce in vivo cellular microenvironment which provide a epithelial-mesenchymal interaction. Because conventional monolayer culture lacks epithelial-mensenchymal interaction, cultivated cells have an morphologic, biochemical, and functional characteristics differ from in vivo tissue. Moreover, it's condition is not able to induce cellular differention due to submerged culture condition. Therefore, the aims of this study were to develop and evaualte the in vitro experimental model that maintains epithelial-mesenchymal interaction by organotypic raft culture, and to characterize biologic properties of three-dimensionally reconstituted oral keratinocytes by histological and immunohistochemical analysis. The results were as follow; 1. Gingival keratinocytes reconstituted by three-dimensional organotypic culture revealed similar morphologic characteristics to biopsied patient specimen showing stratification, hyperkeratinosis, matutation of epithelial architecture. 2. Connective tissue structure was matured, and there is no difference during stratification period of epithelial 3-dimensional culture. 3. The longer of air-exposure culture on three-dimensionally reconstituted cells, the more epithelial maturation, increased epithelial thickness and surface keratinization 4. In reconstitued mucosa, the whole epidermis was positively stained by anti-involucrin antibody, and there is no difference according to air-exposured culture period. 5. The Hsp was expressed in the epithelial layer of three-dimensionally cultured cells, especially basal layer of epidermis. The change of Hsp expression was not significant by culture stratification. 6. Connexin 43, marker of cell-cell communication was revealed mild immunodeposition in reconstitued epithelium, and there is no significant expression change during stratification. These results suggest that three-dimensional oragnotypic co-culture of normal gingival keratinocytes with dermal equivalent consisting type I collagen and gingival fibroblasts results in similar morphologic and immunohistochemical characteristics to in vivo patient specimens. And this culture system seems to provide adequate micro-environment for in vitro tissue reconstitution. Therefore, further study will be focused to study of in vitro gingivitis model, development of novel perioodntal disease therapeutics and epithelial-mensenchymal interaction.
Regeneration of the periodontal tissue destroyed by periodontal disease is one of the final goals of periodontal therapy. In the past few years, periodontists have used various alloplastic grafting materials in an attempt to regenerate bone lost from periodontal disease. These materials have used widely because they have shown to be nontoxic, biologically compatible with surrounding host tissue and chemically similar to bone. The purpose of this study was to investigate the effect of Porous Resorbable Calcium Carbonate and Porous Replamineform Hydroxyapatite on the regeneration of the alveolar bone and the healing of roots transplanted into the periodontally diseased extraction sockets of dogs. The experimental chronic periodontitis was induced by elastic ligatures on the 2nd and 3rd mandibular premolars of 2 adult dogs for 8weeks after surgically creating periodontal defect. The extracted root were split in half along the long-axis, and the extend of plaque exposure was marked on the root surfaces with burs. The roots were inserted in extraction sockets with Porous Resorbable Calcium Carbonate(PRCC) in left side and with Porous Replaminefrom Hydroxyapatite(PRH) in right side. The flaps were sutured to cover the sockets completely. The animals were sacrificed after 12 weeks of healing, and the specimens were examined histologically. The results were as follows: 1. No inflammatory reactions were observed in either groups. 2. Hoot resorption was observed in both groups while the general outline of the roots were maintained. 3. PRCC was almost completely resorbed and replaced with new bone, while R.H.A. was not resorbed & remained encased in newly-formed C-T and alveolar bone. 4. PRH was encapsulated with alveolar bone which has been deposited from apical & lateral area of the sockets, while the coronal portion of the sockets were filled with C-T. 5. In both groups, the resorbed portions of the roots were replaced with new bone. These results suggest that either PRCC or PRH may not interfere with bone formation or healing in extraction sockets, and in some degree, retard the root resorption. Because the roots maintained in anatomy, we think that graft materials prevent the root resorption.
Regeneration of periodontal tissue after a loss of attachment due to disease or trauma repesents an important issue in dentistry, and various bone graft materials have been used to regenerated lost periodontal tissue and restore proper fuctions. Among those, allografts have been extensively researched and widely used clinically, since they are known to possess an excellent osteoinduction capability and result in proper topography of alveolar bone. Regeneration of periodontal tissue in supraalveolar defects may be technically difficult. However, a large amount of regeneration has been observed by complete tissue coverage of involved teeth. In this study, supraalveolar defects in adult dogs were treated with periodontal surgery, decalcified freez-dried bone allograft, complete tissue coverage was attained, and effects on repair and regeneration of alveolar bone, cementum and periodontal ligament were studied. Exposure of premolar furcation of adult dogs was attained by removing marginal alveolar bone down to 5mm from CEJ, and root surfaces were planed with curettes. On the left side, defects were treated without any allograft(Control Group). On the right side, a DFDB was used(Experimental Group). In all groups, flaps were coronally positioned and sutured, completely submerging the treated defects. At two weeks, the crown were exposed 2-3mm. Healing progresses were histologically observed after eight weeks and the results were as follows : 1. Distance from CEJ to AJE was : $2.82{\pm}0.66mm$ in the control group, $1.71{\pm}0.51mm$ in experimental group, with significant differences between groups.(P<0.01) 2. Periodontal repair was : $2.18{\pm}0.66mm$ in the control group, $3.29{\pm}0.51mm$ in experimental group, with significant differences between groups.(P<0.01) 3. Connective tissue repair was : $1.43{\pm}0.52mm$ in the control group, $0.76{\pm}0.47mm$ in experimental group, with significant differences between groups.(P<0.01) Orientation of connective tissue fibers in relation to root surfaces was : mostly parallel in the control group, vertical or parallel or irregular in experimental group. 4. The amount of cementum formation was : $1.66{\pm}0.58mm$ in the control group, $2.86{\pm}0.66mm$ in experimental group, with significant differences between groups. 5. The amount of alveolar bone formation was : $0.76{\pm}0.72mm$ in the control group, $2.53{\pm}0.56mm$ in experimental group, with significant differences between groups.(P<0.01)
Purpose: Biphasic calcium phosphates have been of great interest recently. Mixing adequate ratios of hydroxyapatite(HA) and beta-tricalcium phosphate($\beta$-TCP) allowed to control the resorption rate without distorting its osteoconductive property. This study evaluated the bone formation effect of newly developed biphasic calcium phosphate(BCP) in calvarial defect of rabbits. Materials and Methods: 6 male New Zealand rabbits were used. Four defects with 8mm in diameter were created on each animal. BCP with HA/$\beta$-TCP ratio of 7:3 and particle size of $0.5{\sim}1.0\;mm$ was used as the test group and bovine bone with $0.25{\sim}1.0\;mm$ particle size, as the control group. Both test and control group materials were randomly implanted in the calvarial defects and were covered witha polymer membrane. The animals were sacrificed after 12, 24, and 48 weeks of implantation under general euthanasia. Resin blocks were obtained and were stained by masson's trichrome for histological observation. Results: Overall results were uneventful without any defect exposure or inflammation. The amount of new bone formation and bone maturity increased with increase in healing period at both groups. New bone in test group was mostly formed along the material particle surrounded by osteoblasts, and observation of osteoblastic stream was also present. Bone maturity increased as it was closer to thedefect margins. Under the same healing period, the test group showed more bone formation than the control group with more stable bovine bone particles remaining even after 48 weeks, whereas considerable resorption took place in BCP. Almost total defect closure was observed in test group with new bone formation in the central part of the defect. However, limited new bone formation was observed in the control group. Conclusion: Within the limits of the study, the present study reveals the newly developed BCP to be a good osteoconductive material. However, further studies are needed to be conducted in a different study model with a larger sample size.
The purpose of this study was to evaluate the structural change of root surface and the occlusion of dentinal tubule following $CO_2$ laser treatment. Seven extracted healthy human premolar werw curetted, sectioned, and four specimens were randomly assigned to each of 6 different treatment groups : 1) untreated EDTA etched control: 2) root plande only: 3) $CO_2$ laser treated with 2W mode 6(10msec/pulse, 20pps) for 1 minute: 4) $CO_2$ laser treated with 2W mode 6(lOmsec/pulse, 20pps) for 2 minutes: 5) $CO_2$ laser treated with 2W mode 7(20msec/pulse, 20pps) for 1 minute: 6) $CO_2$ laser treated with 2W mode 7(20msec/pulse, 20pps) for 2 minutes. Following the prescribed treatment, the specimens were prepared for SEM evaluation. Results showed that $CO_2$ laser may be effective to occlude dentinal tubules tor dentin sensitivity treatment. The effect of dentinal tubule occlusion was enhanced with increasing the total energy level lased to specimen regardless of lasing mode. The structural changes of root surfaces were restricted to superficies, and these changes included fissuring, charring, crater formation over the smooth lava like texture. The charring and crater formation implying root damage was observed in the case of the longer duration of a pulse. The results of the present study suggests that the pulsed $CO_2$ laser with shorter pulse duration and longer exposure time can be used effectively in order to obtain the optimal dentinal tubule occlusion with minimal root damage.
The purpose of this study was to evaluate the in vitro effects of Nd:YAG laser irradiation on removal of a root surface smear layer after root planing in comparison with Tetracycline HCl. The 60 extracted human teeth due to severe periodontal disease were vigorously scaled and root planed with Gracey curet. Thirty specimen($5{\times}5{\times}2mm$) were obtained from root planed surface of 30 human teeth and assigned randomly to one of three groups : root planed group(5 specimen), Tetracycline HCI group(5 specimen, burnished for 5 minutes), and Nd:YAG laser group(25 specimen, German Dental Laser, Fotona Twinlight). Nd:YAG laser group was divided into 4 subgroups according to power of 1W, 1.5W, 2W, 3W at frequency to 10Hz. The specimen were then fixed, and examed by Scanning electron microscopic study. 30 of 60 human teeth used to measurement of the intrapulpal temperature rise during laser irradiation. Laser-irradiated surface exhibited various surface texture from relative flat surface to irregular surface with patent dentinal tubules of various shape and size. In some area, the root surface alteration which are carbonization, pit and crater formation and melting and resolidification were observed. The number of exposed dentinal tubules per unit($100_{\mu}m^2$) on tetracycline HCI group was more than that in the laser group below 1.5W of power(150mJ/pulse) and was significantly less than that in laser group above 2W of power(200mJ/pulse)(P<0.OOl). As power increased the intrapulpal temperature rise also increased. The result suggested that the parameter which effectively remove root surface smear layer than tetracycline HCI may cause thermal damage to pulp and root surface alteration result from laser exposure would indicate need for additional instrumentation. Thus, Nd:YAG laser irradiation in these parameter may not be appropriate for clinical use as adjunct to conventional periodontal therapy.
The purpose of this study was to evaluate exophytically vertical bone formation in residual ridge of the beagle dog by the concept of guided bone regeneration with a titanium reinforced e-PTFE membrane combined with irradiated cancellous human bone. Twelve male beagle dogs(mean age 1.5 years and mean weight 12kg) were used for this study. The alveolar ridges after extraction of all mandibular premolars were surgically and horizontally removed. At 8 weeks after extractions, full-thickness flap was reflected and cortical bone was removed with round bur and copious irrigation. Rectangular parallelepiped(10mm in length, 5mm in width, and 4mm in height) bended with titanium-reinforced e-PTFE(TR e-PTFE) membrane was placed on the decorticated alveolar ridge, fixed with metal pins and covered with full-thickness flap and assigned as a control group. Test groups ere treated with TR e-PTFE membrane filled with irradiated cancellous human bone. Of twelve beagle dogs, four control dogs and four test dogs without membrane exposure to oral cavity were sacrificed at 8 and 16 weeks respectively. The surgical sites were dissected out, fixed in 4% buffered formaldehyde, dyed using a Villanueva staining technique, and processed for embedding in plastic resin. The cutting and grinding methods were routinely processed for histologic and histomophometric analyis of exophytic bone formation as well as statistical analysis. The results of this study were as follows: 1. Exophytic bone formation in the both of experimental groups was increased respectively after surgery from 23.40% at 8 weeks to 46.26% at 16 weeks in the control groups, from 40.23% at 8 weeks to 47.11% at 16 weeks in the test groups(p<0.05). 2. At 8 weeks after surgery, exophytic bone formation was made 40.23% in the test groups and 33.40% in the control groups. Exophytic bone formation was significantly made in the test group more than in the control group. At 16 weeks after surgery, exophytic bone formation was made 44.11% in the test groups and 46.26% in the control groups. Exophytic bone formation was made in the test groups more than in the control groups, but there was no statistically significant differences. 3. The membrane was fixed with metal pins to closely contact it to the bone surface. So, collapse and deviation of the membrane could be prevented and in growth of connective tissue also could be blocked from the periphery of the membrane. On the basis of these findings, wee suggest that intraoral experimental model for exophytic bone formation may be effective to evaluate the effect of bone graft material. And it indicates that combined use of membrane and ICB graft material is more effective than use of membrane only for exophytic bone formation.
파노라마 X-ray (PX) 및 Cone Beam Computed Tomography (CBCT)와 같은 구강 영상 기술은 영상 촬영 시 환자의 편의성과 전체 치아 정보를 시각화할 수 있는 능력으로 인해 치과 진료소에서 가장 선호되는 영상 기법이다. PX는 일상적인 임상 치료에 선호되고, CBCT는 복잡한 수술 및 임플란트 치료에 선호된다. 그러나 PX는 3차원 공간정보가 부족하다는 한계가 있는 반면 CBCT는 환자에게 높은 방사선 노출을 초래한다. PX가 이미 사용 가능한 경우 PX로부터 3D강 구강구조를 복원함으로써 추가 비용을 줄이고 방사선량을 피할 수 있다. 본 논문에서는 PX 이미지로부터 구강구조의 3차원 복원을 위한 U-Net 기반 딥러닝 프레임워크인 3DentAI를 제안한다. 제안된 프레임워크는 PX 이미지에서 깊이를 추정하기 위한 Attention U-Net 기반 재구성 모듈, 사전 정의된 초점 골 및 광선 데이터를 사용하여 예측된 편평 볼륨을 턱 모양에 정렬하기 위한 재정렬 모듈과, 구강의 원활한 표현을 얻기 위해 누락된 정보를 보간하는 3D U-Net 기반 개선 모듈의 세 가지 모듈로 구성된다. 네트워크를 훈련하기 위해, 쌍을 이루는 PX 및 CBCT 데이터셋 대신에 광선 추적 및 렌더링을 통해 CBCT로 부터 합성한 PX 데이터를 사용하였다. 600명의 환자로 구성된 다양한 데이터셋으로 모델을 훈련한 결과, 낮은 계산 복잡도에도 GAN 기반 모델에 비해 우수한 성능을 보였다.
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