Background: 4-Hexylresorcinol (4HR) is able to increase angiogenesis. However, its molecular mechanism in the human endothelial cells has not been clarified. Methods: As endothelial cells are important in angiogenesis, we treated the human umbilical vein endothelial cells (HUVECs) with 4HR and investigated protein expressional changes by immunoprecipitation high-performance liquid chromatography (IP-HPLC) using 96 antisera. Results: Here, we found that 4HR upregulated transforming growth factor-β (TGF-β)/SMAD/vascular endothelial growth factor (VEGF) signaling, RAF-B/ERK and p38 signaling, and M2 macrophage polarization pathways. 4HR also increased expression of caspases and subsequent cellular apoptosis. Mechanistically, 4HR increased TGF-β1 production and subsequent activation of SMADs/VEGFs, RAF-B/ERK and p38 signaling, and M2 macrophage polarization. Conclusion: Collectively, 4HR activates TGF-β/SMAD/VEGF signaling in endothelial cells and induced vascular regeneration and remodeling for wound healing.
피부재생을 위한 생체재료는 염증반응이 최소화되는 안정한 소재로 빠른 피부재생을 돕기 위해 우수한 생체활성과 생체친화성을 가져야 하며, 세포의 부착과 성장을 돕는 미세구조와 다공성이 있어야 한다. 본 연구에서는 새로운 콜라겐 원으로서의 축산부산물인 오리발을 사용하여 콜라겐을 추출하였고 이를 탈미네랄화된 골분demineralized bone powder, DBP), 돼지 소장점막하 조직(small intestinal submucosa, SIS)과 비교하기 위해 스펀지 형태로 제작하였다. 지지체의 물리, 화학적 특징은 SEM, FTIR을 통해 확인하였다. 세포를 파종하여 MTT를 통해 세포의 부착 및 증식률을 측정하였고, 전염증성 사이토카인의 발현도를 보기 위해 RT-PCR을 실시하였다. 또한 항산화 활성능력을 보기 위해 1,1-diphenyl-2-picrylhydrazyl(DPPH)를 측정하였다. 그 결과 오리발 콜라겐 지지체가 물리적 특성이 우수하고 생체적합성이며, 상처 치유제로서의 가능성을 보여주었다.
The ultimate objective of periodontal treatment is to stop disease progression and to regenerate destroyed periodontal tissues and thereby regain normal function. Growth factors are naturally found polypetides which stimulate many cellular activities pertaining to wound healing by acting as signal molecule in controlling cell movement, proliferation, and matrix production. Platelet derived growth factor (PDGF) is 28,000-35,000 Da molecular weight dimeric protein with 2 long positively charged polypeptide chains connected by sulfide bonds. The purpose of this study is to evaluate histologically the initial guided tissue regeneration in a periodontal defect f a beagle dog treated with a biodegradable membrane formed with polylactic acid (poly-L-lactic acid) and polyglycolic acid loaded with 200ng/$cm^2$ platelet derived growth factor. 2 beagle dogs were used in he experiment. $5mm{\times}6mm$ alveolar bone defect was formed in upper and lower canines and third premolars and a reference notch was placed. PDGF-BB non-containing membrane was used as control. Each defect was randomly assigned to the test roup or the control group. The dogs were sacrificed 3 weeks after membrane placement. Toluidine blue and multiple staining was done for histological analysis. In the 3 week specimen in the control group, no new one formation could be seen. Small amount f bone resorption below the notch could be seen. In the notch, loose connective tissue with infiltration of inflammatory cells could be seen. Also thin discontinuous new cementum could be seen and the membrane still retained its structure. Where PDGF-BB containing membrane was used, new bone formation could be seen in the notch at weeks and also continuous thin cementum could be seen. PDL cells were observed between new bone and new cementum and some were attached to bone and cementum. These results suggest that new bone and cementum formation seen when PDGF-BB loaded membrane was used was due to inhibition of downgrowth of epithelial cells and also due to continuous release of the growth factor. Further study on the resorption characteristics of the membrane nd the release characteristics of the PDGF-BB is necessary. Also, development of a membrane easier to use clinically is necessary.
본 연구는 수증기 증류법을 이용하여 추출한 산국화 유래 hydrosol의 피부각질형성세포(HaCaTs) 증식 및 이주 유도 활성 평가를 통해 피부재상피화 및 상처치유 활성을 확인하였다. 산국화 hydrosol은 HaCaTs의 증식과 이주를 농도 의존적으로 유도하였으며, 특히 $1{\mu}g/mL$에서 음성 대조군(control)에 비해 $143.71{\pm}3.37%$의 증식과, $139.98{\pm}5.72%$의 이주 유도 활성을 나타내었다. 또한, 산국화 hydrosol은 extracellular signal-regulated kinase 1/2 (Erk 1/2)와 serine/threonine-specific protein kinase (Akt)의 인산화를 유의하게 증가시켰을 뿐만 아니라 collagen sprout out growth를 농도 의존적으로 유도하였다. 이러한 결과들을 통해서 산국화 hydrosol은 정상적인 재상피화 과정과 상처치유 등의 활성이 있음을 확인하였고, 향후 화장품 소재로서 응용가능성을 검증하였다.
Purpose: This study is performed to determine the effects of titanium cap with various sizes of pores on bone formation during guided bone regeneration (GBR). Methods: Calvaria from 10 adult male rabbits were chosen as the recipient sites. A trephine bur with a diameter of 10 mm was used to form one round groove on each side of sagittal suture of the cranium, and a round bur with a diameter of 1.5 mm was used to form 6 small holes on the inner circles of round grooves to induce bleeding. In the control group, bone graft was not conducted, and closed titanium cap was fixed in the round groove. Bone graft was not performed in groups 1 and 2, but fixed on titanium caps with 0.2 mm, and 0.5 mm sized pores, respectively. For groups 3, 4, and 5, a synthetic bone graft material (${\beta}$-tricalcium phosphate, Cerasorb$^{(R)}$, Germany) was transplanted, and titanium caps without pore, with 0.2 mm and 0.5 mm sized pore were fixed, respectively. The animals were sacrificed 4 weeks after, and clinical, radiographical, and histomorphometrical evaluation of bone regeneration was performed. Results: In all groups, there were no clinical signs of infection, inflammation or wound dehiscence. Radiographic evaluation revealed well-defined semi-circular radiopacity inside the titanium cap of groups 3, 4, and 5. Histologically, the inner surface of the hemisphere was evenly lined with newly formed bone tissue, as well as grafted bone material in the group 3. In groups 4 and 5, the insertion of connective tissue was observed along the inner surface. However, the overall surface area between the grafts with different holes yielded no statistical significance in the histomorphometrical evaluation. Conclusion: Although the total area of newly formed bone showed no significant difference, excellent bone formation tendency was observed histologically when closed caps were used with bone graft was accompanied.
New techniques for regenerating the destructed periodontal tissue have been studied for many years. Current acceptable methods of promoting periodontal regeneration are basis of removal of diseased soft tissue, root treatment, guided tissue regeneration, graft materials, and biological mediators. Platelet Rich Plasma has been reported as a biological mediator which regulates activities of wound healing progress including cell proliferation, migration, and metabolism. The purpose of this study is to evaluate the effects of using the Platelet Rich Plasma as a regeneration promoting agent for furcation involvement defect. Five adult beagle dogs were used in this experiment. The dogs were anesthetized with Ketamin HCl(0.1 ml/kg, IV)and Xylazine hydrochloride($Rompun^{(R)}$, Bayer, 0.1 ml/kg, IM) and conventional periodontal prophylaxis were performed with ultrasonic scaler and hand instruments. With intrasulcular and crestal incision, mucoperiosteal flap was elevated. Following decortication with 1/2 high speed round bur, degree II furcation defect was made on mandibular third(P3), forth(P4) and fifth(P5) premolar, and stopping was inserted. After 4 weeks, stopping was removed, and bone graft was performed. Ca-P was grafted in P3(experimental group I), Combination of Ca-P and plasma rich platelet were grafted in P4(experimental group II), and P5 was remained at control group.Systemic antibiotics(gentamicin sulfate)and anlgesics(phenyl butazone) were administrated intramuscular for 2 weeks after surgery. Irrigation with 0.1% Chlorhexidine Gluconate around operate sites was performed during the whole experimental period except one day immediate after surgery. Soft diets were fed through the whole experiment period. After 4, 8 weeks, the animals were sacrificed by perfusion technique. Tissue block was excised including the tooth and prepared for light microscope with Gomori's trichrome staining. At 4 weeks after surgery, there were rapid osteogenesis phenomenon on the defected area of the Platelet Rich Plasma plus Ca-P BBP group and early trabeculation pattern was made with new osteoid tissue produced by activated osteoblast. Bone formation was almost completed to the fornix of furcation by 8 weeks after surgery. In conclusion, Platelet Rich Plasma can promote rapid osteogenesis during healing of periodontalregeneration.
The authors have transplanted periodontally involved roots which had been root planed into healthy and periodontally involved extraction sockets, and studied the root resorption patterns as well as its effect on new bone formation and wound healing. Alveolar bone around mandibular premolars of 6 adult dogs has been surgically removed, followed by ligation of orthodontic elastic wires for 8 weeks inducing chronic periodontal disease. After removing the crown portions, roots were extracted, and notches were made on the root surfaces discriminating healthy and periodontally involved areas using burs. Controls and experimental groups were divided as follows. Control I : Transplantation of periodontally involved root into healthy extraction sockets. Control II : Transplantation of periodontally involved root into diseased extraction sockets. Experimental group I : Transplantation of root planed roots into healthy extraction sockets. Experimental group II : Transplantation of root planed roots into diseased extraction sockets. Extraction sockets were sutured after transplantations, completely submerging the roots. Healing progress was histologically observed at 2nd, 8th, 12th, and 20th weeks, and the results were as follows ; 1. No inflammation or infection within the extraction sockets had been observed in all groups throughout the experimental period. 2. Reversal lines were observed at week 2 in all groups, clearly discriminating socket walls and new bone, and numerous blood vessels were observed in the new bone trabeculae. 3. Experimental groups showed markedly less root resorption compared to the controls at week 2, but as time progressed, severe resorptions were present in all groups. 4. Localized areas of new bone ankylosis were observed, and the rest of the areas showed collagen fiber insertion with new bone formation at its periphery. 5. No clear differences were found in healing and alveolar bone regeneration between healthy and diseased extraction sockets. 6. The amount of root resorption and ankylosis had increased up to week 8 and 12, showing ankylosis of new bone and the roots. However, no further increase in ankylosis was observed at week 20. 7. Most of the cementum on healthy roots was directly ankylosed to new bone at week, 2, and were gradually resorbed and replaced by new bone thereafter. These results appear to indicate that root planing may inhibit early root resorption of transplanted roots, but gradual replacement by alveolar bone and collagen fibers eventually occur. Condition of the roots or presence of disease in extraction sockets do not appear to make marked differences in alveolar bone regeneration process.
Objective: This study aimed to ascertain the curative effects of Gamisipjeon-tang (GST) used for wound healing on the skin regeneration of deep second degree burns in mice. Material & Methods: In vitro, the $I{\kappa}B$ kinase (IKK) mRNA expression, inducible nitric oxide synthase (iNOS) mRNA expression, and cyclooxygenase-2 (COX-2) mRNA expression in the GST concentration from 1 mg/$m{\ell}$ to 10 mg/$m{\ell}$ were measured. In vivo, the mice were divided into four groups : the normal group, the BE group (burn-elicited group, control group), the DC group (Duoderm CGF-treated group after burn elicitation), and the GST group (Gamisipjeon-tang treated group after burn elicitation). To determine the anti-inflammatory effects, nuclear factor (NF)-${\kappa}B$ p65, iNOS, COX-2 positive reaction were measured by immunohistochemistry. To estimate the skin regenerative effects, change of burn area, 5-bromo-2'-deoxyuridine (BrdU), and fibroblast growth factor (FGF) positive reaction were analyzed. Results: In vitro, the iNOS, IKK, COX-2 mRNA expression decreased according to the increase of GST concentration. The significant decrease of COX-2, iNOS, NF-${\kappa}B$ positive reaction were the highest in the GST group, followed by the DC group and the BE group (p<0.05). The diameter of burn area was significantly decreased in the GST group as compared to that in the DC and BE group (p<0.05). The BrdU and FGF positive reaction increased more significantly in the GST group than in the DC group, and more significantly in the DC group than in the BE group on the 3rd and 7th day after burn (p<0.05). FGF positive reaction increased in the BE and DC group, whereas it decreased significantly in the GST group on the 14th day (p<0.05). The BrdU positive reaction increased in the BE group, whereas it decreased significantly in the DC and GST group on the 14th day (p<0.05). Conclusions: This study shows that GST could decrease the inflammatory response and accelerate the skin regeneration as compared to the duoderm CGF in mice with deep second degree burns.
This study was performed to evaluate the effects of extracts of Drynariae Rhizoma on the characteristics of rat calvaria cells(RCV) and bone marrow cells(RBM) which have the important role on the bone formation in vitro. Drynariae Rhizoma has been known as the useful herbal medicament for treatment of the wound healing including regeneration of bone fracture, and also has been used to treat the periodontal lesions, tooth mobility, gingival bleeding and pus discharge via sulcus in Oriental Medicine. In control group, the cells were cultured alone with Dulbeco's Modified Eagle's Medium contained with 10% fetal bovine serum, 100U/ml penicillin, $100{\mu}g/ml$ streptomycin, $0.5{\mu}g/ml$ amphotericin-B. In experimental group, extracts of Drynariae Rhizoma(0.1, 1, 5, 10, $50{\mu}g/ml$) were added into the above culture condition. And then each group was characterized by examing the cell proliferation at 1, 3, 7, 14, 21, 30th day, the amount of total protein synthesis and alkaline phosphatase activity of RCV at 2,4th day and those of RBM at 3, 6th day. And also, the calcified nodule of RCV was examed at 3, 5th day in three goup, control, experimental, culture with the PDGF group. The results were as follow ; 1. Both RCV and RBM cells in Drynariae Rhizoma-treated experimental group proliferated more rapidly than nontreated control group. The experimental group below $5{\mu}g/ml$ Drynariae Rhizoma-treated showed more prominent cell proliferation from the 7th day to the 21st day than the control group and above $10\;{\mu}g/ml$ treated group in RCV. 2. Amount of total protein synthesis was more increased in Drynariae Rhizomatreated group than in control group. In $5{\mu}g/ml$ Drynariae Rhizoma-treated group showed most prominent protein synthesis of the any other exrperimental group and control group. 3. Alkaline phosphatase activity also more increased in Drynariae Rhizomatreated group than control group. 4. Mineralized nodules in Drynariae Rhizoma-treated group were more than not in control group but also in PDGF-treated group. From the above results, Drynariae Rhizoma appeared to enhanced the proliferation, protein synthesis, alkaline phosphatase activity and cellular ability of mineralized nodule formation than PDGF. So that, we conclude that Drynariae Rhizoma enhances the activities of bone cells which have the important role on the periodontal regeneration and optimal application of Drynariae Rhizoma was thought to be useful as the means in bone regeneration.
The purpose of the present study was to evaluate the effect of bone graft materials including deproteinized bovine bone(DBB), demineralized freeze-dried bone(DFDB), freeze-dried bone(FDB) on bone formation in guided bone regeneration using perforated titanium membrane(TM). 16 adult male rabbits(mean BW 2kg) were used in this study and 4 rabbits allotted to each test group. Intramarrow penetration(diameter 6.5mm) was done with round carbide bur on calvaria to promote blood supply and clot formation in the wound area. The test groups were devided into 4 groups as follows: TM only(test 1), TM +DBB(test 2), TM +DFDB(test 3), TM +FDB(test 4). Perforated titanium membrane was contoured in rectangular parallelepiped shape(0.5mm pore diameter, 10mm in one side, 2mm in inner height), filled the each graft material and placed on the decorticated carvaria. Perforated titanium membrane was fixed with resorbable suture materials. The animals were sacrificed at 2, 8 weeks after the surgery. Non-decalcified preparations were routinely processed for histologic analysis. The results of this study were as follows: 1. Perforated titanium membrane was biocompatible. 2. Perforated titanium membrane had capability of maintaining the space during the healing period but invasion of soft tissue through the perforations of titanium membrane decreased the space available for bone formation. 3. In test 1 group without bone graft material, the amount of bone formation and bone maturation was better than other test groups. 4. Among the graft materials, the effect of freeze-dried bone on bone formation was best. 5. In the test groups using deproteinized bovine bone, demineralized freeze-dried bone, bone formation was a little. The spacemaking capability of the membrane may be crucial for bone formation. The combined treatment with the perforated titanium membrane and deproteinized bovine bone or demineralized freeze-dried bone failed to demonstrate any added effect in the bone formation. Minimization of size and numbers of perforations of titanium membrane or use of occlusive titanium membrane might be effective to acquire predictable results in the vertical bone formation.
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