Background Composite grafts are frequently used for facial reconstruction. However, the unpredictability of the results and difficulties with large defects are disadvantages. Adipose-derived stem cells (ADSCs) express several cytokines, and increase the survival of random flaps and fat grafts owing to their angiogenic potential. Methods This study investigated composite graft survival after ADSC injection. Circular chondrocutaneous composite tissues, 2 cm in diameter, from 15 New Zealand white rabbits were used. Thirty ears were randomly divided into 3 groups. In the experimental groups (1 and 2), ADSCs were subcutaneously injected 7 days and immediately before the operation, respectively. Similarly, phosphate-buffered saline was injected in the control group just before surgery in the same manner as in group 2. In all groups, chondrocutaneous composite tissue was elevated, rotated 90 degrees, and repaired in its original position. Skin flow was assessed using laser Doppler 1, 3, 6, 9, and 12 days after surgery. At 1 and 12 days after surgery, the viable area was assessed using digital photography; the rabbits were euthanized, and immunohistochemical staining for CD31 was performed to assess neovascularization. Results The survival of composite grafts increased significantly with the injection of ADSCs (P<0.05). ADSC injection significantly improved neovascularization based on anti-CD31 immunohistochemical analysis and vascular endothelial growth factor expression (P<0.05) in both group 1 and group 2 compared to the control group. No statistically significant differences in graft survival, anti-CD31 neovascularization, or microcirculation were found between groups 1 and 2. Conclusions Treatment with ADSCs improved the composite graft survival, as confirmed by the survival area and histological evaluation. The differences according to the injection timing were not significant.
Park, Jae-Young;Hwang, Woo-Jin;Jeong, Seong-Nyum;Kim, Yun-Sang;Pi, Sung-Hee;You, Hyung-Keun;Shin, Hyung-Shik
Journal of Periodontal and Implant Science
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v.39
no.2
/
pp.167-176
/
2009
Purpose: The purpose of this study is to histologically and histomorphometrically evaluate the effect of PLGA on bone regeneration compared with bone graft material. Methods: The experimental study was conducted in 10 rabbits with 2 different healing periods of 2 and 4 weeks. Following surgical exposure of the calvarium, 4 circular bone defects with a diameter of 4.6mm were formed. Rabbits were divided into control group, test groups I, and II. 10 defects assigned to the test group Ⅰ were grafted with Nu-oss and other 10 defects assigned to the test group II were grafted with PLGA. The rest of the defects were in the negative control group. At 2nd and 4th week after surgery, 10 rabbits were sacrificed through intracardiac perfusion and then specimens were obtained. Histological analysis was performed following staining with trichorme and transversal sectioning of the calvarial bone. Results: A group which used PLGA showed tissue reactions characterized by severe inflammation, rather than distinctive new bone formation. Conclusions: The present experimental investigations have failed to prove any beneficial effects of PLGA. PLGA used in this study exhibited foreign body reactions and a less favorable pattern of new bone formation in comparison to control group. Conclusion: PLGA did not function as scaffold. Further investigations of many types of micro PLGA that could improve its potential in GBR procedures are needed.
To better define the relationship between dermal regeneration and wound contraction and scar formation, the effects of epidermal growth factor (EGF) loaded in collagen sponge matrix on the fibroblast cell proliferation rate and the dermal mechanical strength were investigated. Collagen sponges with acid-soluble fraction of pig skin were prepared and incorporated with EGF at 0, 4, and 8 $\mu$g/1.7 $cm^{2}$. Dermal fibroblasts were cultured to 80$\%$ confluence using DMEM, treated with the samples submerged, and the cell viability was estimated using MTT assay. A deep, $2^{nd}$ degree- burn of diameter 1 cm was prepared on the rabbit ear and the tested dressings were applied twice during the 15-day, post burn period. The processes of re-epithelialization and dermal regeneration were investigated until the complete wound closure day and histological analysis was performed with H-E staining. EGF increased the fibroblast cell proliferation rate. The histology showed well developed, weave-like collagen bundles and fibroblasts in EGF-treated wounds while open wounds showed irregular collagen bundles and impaired fibroblast growth. The breaking strength (944.1 $\pm$ 35.6 vs. 411.5 $\pm$ 57.0 Fmax, $gmm^{-2}$) and skin resilience (11.3 $\pm$ 1.4 vs. 6.5 $\pm$ 0.6 mJ/$mm^{2}$) were significantly increased with EGFtreated wounds as compared with open wounds, suggesting that EGF enhanced the dermal matrix formation and improved the wound mechanical strength. In conclusion, EGF-improved dermal matrix formation is related with a lower wound contraction rate. The impaired dermal regeneration observed in the open wounds could contribute to the formation of wound contraction and scar tissue development. An extraneous supply of EGF in the collagen dressing on deep, $2^{nd}$ degree-burns enhanced the dermal matrix formation.
Kim, Yong-Yun;Kwon, Kyung-Hwan;Choi, Moon-Ki;Oh, Sung-Hwan;Min, Seung-Ki
Maxillofacial Plastic and Reconstructive Surgery
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v.27
no.2
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pp.140-150
/
2005
Maxillary sinus lifting procedure and bone grafting are used to reconstruct atrophic maxillae. These procedure are usually followed by the placement of endosseous dental implants. Different materials and techniques can be used for sinus bone grafting. Platelets are known to contain various growth factors involved in the repair of the vasculature and tissues, and it is known that the specialized platelet secretory granules, the alpha granules, contain platelet derived growth factor(PDGF), transforming growth factor-beta(TGF-beta), insuline like growth factor-I(IGF-I), epidermoid growth factor(EGF), and others. This study was to evaluate the effect of PRP on bone formation in a sinus bone grafting. Twelve rabbits were included in this randomized, blinded, prospective pilot study. In experimental group, sinus bone grafting with autobone and platelet rich plasma. In control group, sinus bone grafting with only autobone. Rabbits were sacrificed at 2nd, 4th, 8th, 12th weeks postoperatively. Clinical and radiographic tests, histological analysis were conducted to compare both sides. In clinical examination, there in no significant difference between experimental group and control group. But, in radiographic examination, a distinct incresed in the radiopaque of the PRP experimental group at 2nd and 4th weeks. The histologic examination revealed that more new bone formation and osteoblast activity were seen in experimental group at 2nd and 4th weeks. In conclusion, PRPs action in sinus bone grafting had a capacity of increased new bone formation in a early bone healing stage.
Purpose: The aim of this study was to investigate the efficacy of photo-crosslinked gelatin methacryloyl (GelMa) hydrogel containing calcium phosphate nanoparticles (CNp) when applying different fabrication methods for bone regeneration. Methods: Four circular defects were created in the calvaria of 10 rabbits. Each defect was randomly allocated to the following study groups: 1) the sham control group, 2) the GelMa group (defect filled with crosslinked GelMa hydrogel), 3) the CNp-GelMa group (GelMa hydrogel crosslinked with nanoparticles), and 4) the CNp+GelMa group (crosslinked GelMa loaded with nanoparticles). At 2, 4, and 8 weeks, samples were harvested, and histological and micro-computed tomography analyses were performed. Results: Histomorphometric analysis showed that the CNp-GelMa and CNp+GelMa groups at 2 weeks had significantly greater total augmented areas than the control group (P<0.05). The greatest new bone area was observed in the CNp-GelMa group, but without statistical significance (P>0.05). Crosslinked GelMa hydrogel with nanoparticles exhibited good biocompatibility with a minimal inflammatory reaction. Conclusions: There was no difference in the efficacy of bone regeneration according to the synthesized method of photo-crosslinked GelMa hydrogel with nanoparticles. However, these materials could remain within a bone defect up to 2 weeks and showed good biocompatibility with little inflammatory response. Further improvement in mechanical properties and resistance to enzymatic degradation would be needed for the clinical application.
The aim of the study was to assess the in vitro effect of 808-nm InGaAs diode laser on rabbit articular chondrocyte proliferation and sulphated glycosaminoglycan (sGAG) synthesis in alginate bead. Previous studies revealed either positive or negative stimulatory effects of laser on different types of cells. A 808-nm InGaAs diode laser at 1.0W power output was used to irradiate the rabbit chondrocytes in alginate beads with energy densities of $31J/cm^2$ (G 1) and $62J/cm^2$ (G 2) corresponding to the experimental groups for 10 seconds and 20 seconds, respectively at 24, 48, 72 and 96 hours after seeding. Control group was left untreated. MTT assay was performed at 1 week and 2 weeks after the $1^{st}$ laser irradiation in alginate beads. sGAG synthesis in alginate beads at 1 week and 2 weeks were determined by DMMB assay. Histological evaluation for cellular distribution and sGAG deposition around the cells were performed by alcian blue stain. MTT assay revealed no positive stimulatory effect in cell proliferation in alginate bead. DMMB assay results showed significantly increased sGAG production in G 2 chondrocytes at 2 weeks. Image analysis of alcian blue stained slides also showed significantly higher percentage of positive alcian blue stain in G 2 chondrocytes. This result suggests that 808-nm InGaAs diode laser with 1.0 W power output although cannot stimulate cell proliferation it can increase the cell secretion activity and sGAG deposition in alginate beads.
Objective : Endovascular mechanical thrombectomy (MT) has been regarded as one of the standard treatments for acute ischemic stroke caused by large vessel occlusion. Despite the wide use of stent retrievers for MT, arterial intimal damage caused when deployed stent is pulled has been a certain disadvantage. We hypothesized that statin could protect and stabilize vessel damage after endovascular MT using a stent retriever. In this animal study, we observed the protective effects of the statins towards MT-induced vessel wall injury. Methods : Twenty-eight carotid arteries of fourteen rabbits were used in the experiments with MT using stent retriever. We divided the rabbits into four groups as follows : group 1, negative control; group 2, positive control; group 3, statin before MT; and group 4, statin after MT. After MT procedures, we harvested the carotid arteries and performed histomorphological and immunohistochemical analyses. Results : In histomorphological analysis with hematoxylin and eosin and Masson's trichrome stain, significant intimal thickening (p<0.05) was observed in the positive control (group 2), compared to in the negative control (group 1). Intimal thickening was improved in the statin-administered groups (groups 3 and 4 vs. group 2, p<0.05). We also observed that statin administration after MT (group 4) resulted in a more effective decrease in intimal thickness than statin administration before MT (group 3) (p<0.05). We performed immunohistochemical analysis with the antibodies for tumor necrosis factor-alpha (TNF-α), cluster of differentiation (CD)11b, and CD163. In contrast to the negative control (group 1), the stained percentage areas of all immunological markers were markedly increased in the positive control (group 2) (p<0.05). Based on statin administration, the percentage area of TNF-α staining was significantly reduced (p<0.05) in group 3, compared to the positive control group (group 2). However, significant differences were not observed for CD11b and CD163 staining. In group 4, no significant differences were observed for TNF-α, CD11b, and CD163 staining (p≥0.05). The differences in the percentage areas of the different markers between the statin-administered groups (groups 3 and 4) were also not revealed. Conclusion : We presented that statin administration before and after MT exerted protective effects towards vessel wall injury. The efficacy of statins was greater post-administration than pre-administration. Thus, statin administration in routine prescriptions in the peri-procedural period is strongly advised.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.34
no.3
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pp.250-265
/
2008
Distraction osteogenesis is a well-established clinical treatment for limb length discrepancy and skeletal deformities. Appropriate mechanical tension-stress is believed not to break the callus but rather to stimulate osteogenesis. In contrast to fracture healing, the mode of bone formation in distraction osteogenesis is primarily intramembranous ossification. Although the biomechanical, histological, and ultrastructural changes associated with distraction osteogenesis have been widely described, the basic biology of the process is still not well known. Moreover, the molecular mechanisms in distraction osteogenesis remain largely unclear. Recent studies have implicated the growth factor cascade is likely to play an important role in distraction. And current reserch suggested that mechanical tension-stress modulates cell shape and phenotype, and stimulates the expression of the mRNA for bone matrix proteins. The purpose of this study is to examine the pattern of expression of growth factors($TGF-{\beta}1$, IGF-I, bFGF) and extracellular matrix proteins(osteoclacin, osteonectin) related to osteogenesis by osteodistraction of the mandible in rabbits. 24 rabbits is used for this experiment. Experimental group are gradual distraction(0.7mm, twice/day), acute distraction(1.4mm, twice/day) and control group is only osteotomized. After 5 days latency, osteotomic site is distracted for each 7 days and 3.5 days. Consolidation period is 28 days. The animal is sacrificed at the 3th, 7th, 14th, 28th. The distracted bone is examined by immunohistochemical analysis and RT-PCR analysis. The results obtained from this study were as follow : No significant difference was found on clinical examination according to distraction rate, but gradual distraction was shown to improve regenerate bone formation on radiographic and histologic examination. Growth factors and extracelluar matrix proteins expression increased in distraction group than control group. From these results, it could be stated that graudal distraction is shown to improve and accelerate bone formation and mechanical stress like distraction has considerable effects on osteogenesis related factors. And rabbit is the most appropriate animal model for further reseach on the molecular mechanisms that mediate osteodistraction. It is believed that understanding the biomolecular mechanisms that mediate distraction osteogenesis may guide the development of targeted strategies designed to improve distraction osteogenesis and accelerate bone healing.
Purpose: This study was conducted in order to compare the efficacy of osseointegration of three different calcium metaphosphate (CMP) coated implants in the rabbit's femur. Materials and Methods: Twenty four rabbits and three different type of CMP coated implants and RBM implants were used in this study. The animals were divided into 4 groups on the basis of implant surface characteristics. Two implants were installed into the condyle of femur of each rabbits. The animals were sacrificed at 2 and 4 weeks after installation. The undecalcified specimens were prepared for histological, radiological examination and histomorphometric analysis of implant-bone contact ratios (BIC) and bone area ratio (BA). Results: Two implants were failed to osseointegrate and implant success rate was 95.2%. There were not any significant inflammatory response in all groups. Fluorescent image at 4 weeks shows that remodeling is slower in RBM group than CMP group. CMP III showed more active remodeling than CMP I, II. In histomorphologic analysis, BIC ratio at 2 weeks was lower than 4 weeks. Conclusion: The results suggest that the ratios of CMP coated implants were higher than that of RBM control group but there is no significantly difference between RBM group and CMP group. In conclusion, CMP coated implant had more clinical availability than RBM implants.
The tendon is a dense connective tissue that connects muscle to bone and plays an essential role in joint motion. The injured tendon heals slowly owing to its low cellularity and vascularity. This study aimed to evaluate and compare the effects of regenerative injection therapy (RIT), 20 % dextrose prolotherapy (DP), and platelet-rich plasma (PRP) injections that can promote tendon healing. Twenty-one New Zealand white rabbits were divided into the control, DP, and PRP treatment groups. The superficial digital flexor tendon (SDFT) of the right hindlimb of each rabbit was used. A round defect of 2 mm was induced. Approximately 0.2 mL of 20% dextrose and autologous PRP were injected into the proximal and distal ends of the SDFT mass. Radiographic and ultrasonographic examination and cross-sectional area (CSA) calculations were performed pre-operatively and at 2, 4, and 8 weeks. The SDFT of both limbs was transected for biomechanical and histomorphometric evaluations. The SDFT of the left limb was transected for intact control. Semi-quantitative analysis was performed to evaluate the histomorphometric properties. Additional analysis was performed using H&E, Masson's trichrome, and immunohistochemical staining. The biomechanical evaluation showed that the treatment groups had higher tensile strength compared to the defect control group, while the PRP group had higher tensile strength than the DP group. On histological examination, the treatment groups appeared to be relatively closer to the remodeling phase of the healing process than the defect control group; the characteristics of the PRP group were closer to the remodeling phase than those of the DP group. The ultrasonographic examination showed different tendencies. Increased values in the CSA were observed during the early period in the treatment groups. This study suggests that PRP and DP can promote the healing of tendon injury, and these effects were superior with PRP than that with DP.
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