• IMMUNE NETWORK
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• v.5 no.1
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• pp.36-44
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• 2005

Background: The anti-tumor therapeutic effect of autologous tumor cell lysate pulseddendritic cells (DCs) was studied for non-immunogenic and immune suppressive lung cancer model. To test the possibility as an adjuvant therapy, minimal residual disease model was considered in mouse in vivo experiments. Methods: Syngeneic 3LL lung cancer cells were inoculated intravenously into the C57BL/6 mouse. Autologous tumor cell (3LL) or allogeneic leukemia cell (WEHI-3) lysate pulsed-DCs were injected twice in two weeks. Intraperitoneal DC injection was started one day (MRD model) after tumor cell inoculation. Two weeks after the final DC injection, tumor formation in the lung and the tumor-specific systemic immunity were observed. Tumor-specific lymphocyte proliferation and the IFN-${\gamma}$ secretion were analyzed for the immune monitoring. Therapeutic DCs were cultured from the bone marrow myeloid lineage cells with GM-CSF and IL-4 for 7 days and pulsed with tumor cell lysate for 18 hrs. Results: Compared to the saline treated group, tumor formation was suppressed in 3LL tumor cell lysate pulsed-DC treated group, while 3LL-specific immune stimulation was minimum. WEHI-3-specific immune stimulation occurred in WEHI-3 lysate-pulsed DC treated group, which had no correlation with tumor regression. Conclusion: The data suggest the possible anti-tumor effect of cultured DCs as an adjuvant therapy for minimal residual disease state of lung cancer. The significance of immune modulation in DC therapy including the possible involvement of NK cell as well as antigen-specific cytotoxic T cell activity induction was discussed.

• Archives of Plastic Surgery
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• v.42 no.2
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• pp.143-149
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• 2015

Background Adipose tissue damage of cryopreserved fat after autologous fat transfer is inevitable in several processes of re-transplantation. This study aims to compare and analyze the survivability of adipocytes after thawing fat cryopreserved at $-20^{\circ}C$ by using thawing methods used in clinics. Methods The survival rates of adipocytes in the following thawing groups were measured: natural thawing at $25^{\circ}C$ for 15 minutes; natural thawing at $25^{\circ}C$ for 5 minutes, followed by rapid thawing at $37^{\circ}C$ in a water bath for 5 minutes; and rapid thawing at $37^{\circ}C$ for 10 minutes in a water bath. The survival rates of adipocytes were assessed by measuring the volume of the fat layer in the top layers separated after centrifugation, counting the number of live adipocytes after staining with trypan blue, and measuring the activity of mitochondria in the adipocytes. Results In the group with rapid thawing for 10 minutes in a water bath, it was observed that the cell count of live adipocytes and the activity of the adipocyte mitochondria were significantly higher than in the other two groups (P<0.05). The volume of the fat layer separated by centrifugation was also measured to be higher, which was, however, not statistically significant. Conclusions It was shown that the survival rate of adipocytes was higher when the frozen fat tissue was thawed rapidly at $37^{\circ}C$. It can thus be concluded that if fats thawed with this method are re-transplanted, the survival rate of cryopreserved fats in transplantation will be improved, and thus, the effect of autologous fat transfer will increase.

• Archives of Plastic Surgery
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• v.45 no.5
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• pp.441-448
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• 2018

Background Previously, surveys have been used to investigate breast reconstruction statistics. Since 2015, breast reconstruction surgery after mastectomy has been covered by the National Health Insurance Service in Korea, and data from breast reconstruction patients are now available from the Health Insurance Review and Assessment Service (HIRA). We investigated statistics in breast reconstruction in Korea through statistics provided by the HIRA Big Data Hub. Methods We investigated the number of cases in mastectomy and breast reconstruction methods from April 1, 2015 to December 31, 2016. Data were furnished by the HIRA Big Data Hub and accessed remotely online. Results were tabulated using SAS Enterprise version 6.1. Results The 31,155 mastectomy cases included 7,088 breast reconstruction cases. Implant-based methods were used in 4,702 cases, and autologous methods in 2,386. The implant-based reconstructions included 1,896 direct-to-implant and 2,806 tissue-expander (2-stage) breast reconstructions. The 2-stage tissue-expander reconstructions included 1,624 expander insertions (first stage) and 1,182 expander-to-permanent-implant exchanges (second stage). Of the autologous breast reconstructions, 705 involved latissimus dorsi muscle flaps, 498 involved pedicled transverse rectus abdominis myocutaneous (TRAM) flaps, and 1,183 involved free-tissue transfer TRAM flaps, including deep inferior epigastric perforator free-tissue transfer flaps. There were 1,707 nipple-areolar complex reconstructions, including 1,565 nipple reconstructions and 142 areola reconstructions. The 1-year mean number of breast reconstructions was 4,050. Conclusions This was the first attempt to evaluate the total number of breast reconstruction procedures using accurate, comprehensive data, and our findings may prove valuable as a foundation for future statistical studies of breast reconstruction procedures in Korea.

• Archives of Plastic Surgery
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• v.33 no.5
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• pp.601-605
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• 2006

Purpose: Large skin defect by various causes, should be covered by autologous skin graft. But, the donor site of autologous skin graft is limited and leaves permanent donor scar and contracture. There have been our trial to engineer artificial skin using allogenic dermis (AlloDerm) with basement membrane. Methods: Dermal and epidermal layer were separated by immersing in dipase solution for 30 minutes, and the separated layers were treated with 0.05% trypsin for 10 minutes. And then each layer was cultivated to fibroblasts and keratinocytes on a culture medium. Fibroblasts were first penetrated into basement membrane of allogenic dermis facing down, then allogenic dermis was flipped over to face up and keratinocytes were transplanted to allogenic dermis. Results: Observing artificial skin fabricated in vitro, we found following: 1) The artificial skin opened in air for 5 days formed epidermal layer. In dermal layer, fibroblast was distributed evenly among all. 2) The artificial skin opened in air for 30 days formed thicker and thicker, and it formed basement membrane, spinous and granular layers. PAS stain to confirm existence of basement membrane showed positive reaction. 3) Cytokeratin 10 stain to confirm the formation of epidermal layer showed positive reaction. 4) The formation of thick keratin, lamellar body and desmosome similar to human skin were observed in result of an electron micrograph. Conclusion: As a result of research, the structure seen in normal skin such as rete ridge, is found in reproduced artificial skin. This type of artificial skin can be used as a useful model for investigating skin disease and for clinical application also.

• Archives of Plastic Surgery
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• v.32 no.4
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• pp.529-532
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• 2005

Expanding cells ex-vivo is very important in tissue-engineering. Culture medium is usually supplemented with fetal bovine serum(FBS) in most of the experiments. However, cells grown in bovine serum media may posses the possibilities of disseminating bovine diseases and/or stimulating the patient's immune reactions. To overcome these problems, autologous or homologous serum should be used instead of the FBS. The purpose of this study is to compare cell proliferation and collagen synthesis depending on the kind of sera mixed on media and to provide a guideline on applying established experimental data to clinical cases. Human dermal fibroblasts were obtained from four patients. Five thousand cells per well in 96-well plates were incubated DMEM/F-12 Nutrient with varying serum mixture; 10% autologous serum, 10% homologous serum, and 10% FBS. Five days after incubation fibroblast proliferation and collagen production were determined by MTT assay and CICP enzyme immunoassay. The mean cell number were; $3.95{\times}10^4/well$, $2.97{\times}10^4/well$ and $2.30{\times}10^4/well$, respectively. The average amounts of collagen synthesized were; 238.13 ng/ml, 204.88 ng/ml, and 163.88 ng/ml in each. These results show that the use of human serum mixture may contribute to, not only preventing disseminated infection of bovine diseases. but also increase cell proliferation and collagen synthesis without simulating the patient's immune reactions.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.35 no.5
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• pp.346-352
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• 2009

There have been reports of successful bone formation with sinus floor elevation by simply elevating the maxillary sinus membrane and filling the sinus cavity below the lifted sinus membrane with a blood clot. But, in a review of the current literature, we found no animal study that substantiated blood clot's ability in this respect. The aim of this study was to investigate the effect of the method of maxillary sinus floor augmentation using the patient's own venous blood in conjunction with a sinus membrane elevation procedure. An implant was placed bilaterally in the maxillary sinus of six adult mongrel dogs so that it protruded 8 mm into the maxillary sinus after sinus membrane elevation. On one side of the maxillary sinus, the resultant space between the membrane and the sinus floor was filled with autologous venous blood retrieved from the dog. On the opposite side, the maxillary sinus was left untreated as a control. The implants were left in place for six months. The mean height of the newly formed bone in the sinus was 3.7 mm on the side without venous blood and 3.5 mm on the side with venous blood (p>0.05). There was no difference between the two sides regarding new bone height in the sinus. Our results indicate that filling the space between the lifted sinus membrane and the sinus floor with venous blood has no effect on bone formation around implants placed in the maxillary sinus cavity.

• IMMUNE NETWORK
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• v.6 no.4
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• pp.179-184
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• 2006

Background: Identification of antigen-specific T cells has yielded valuable information on pathologic process and the disease state. Assays for quantification of inflammatory cytokines or lytic-granule molecules have been generally used to evaluate antigen specific T cell response, however their applicability have been hampered due to the limited source of autologous antigen-presenting target cells (APC). Methods: K562, a leukemic cell line deficient of human leukocyte antigen (HLA), was transfected with a gene encoding HLA-A*02 (K562/ A*02) and its function as stimulator cells in inducing activation of HLA-matched T cells was evaluated by IFN-${\gamma}$ enzyme linked immunospot (ELISPOT) assay. Results: The stable transfectant K562/ A*02 pulsed with HLA- A*02 restricted peptide could specifically induce IFN-${\gamma}$ secretion by CD8+ T cells compared to no detectable secretion by CD4+ T cells. However, CD56+ NK cells secreted IFN-${\gamma}$ in both K562/ A*02 with peptide and without peptide. The number of IFN-${\gamma}$ secreted CD8+ T cells was increased according to the ratio of T cells to K562 and peptide concentration. Formalin-fixed K562/ A*02 showed similar antigen presenting function to live K562/ A*02. Moreover, K562/ A*02 could present antigenicpeptide to not only A*0201 restricted CD8+ T cells but also CD8+ T cells from A*0206 donor. Conclusion: These results suggest that K562/ A*02 could be generally used as target having specificity and negligible background for measuring CD8+ T cell responses and selective use of K562 with responsder matched HLA molecules on its surface as APC may circumvent the limitation of providing HLA-matched autologous target cells.

• Archives of Craniofacial Surgery
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• v.19 no.2
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• pp.94-101
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• 2018

Background: Atrophy of muscle and fat often contributes to temporal hollowing after pterional craniotomy. However, the main cause is from the bony defect. Several methods to prevent temporal hollowing have been introduced, all with specific limitations. Autologous bone grafts are most ideal for cranial defect reconstruction. The authors investigated the effectiveness of bony defect coverage and temporal augmentation using pterional craniotomy bone flap. Methods: This study was conducted in 100 patients who underwent brain tumor excision through pterional approach from 2015 to 2016. Group 1 underwent pterional craniotomy with temporal augmentation and group 2 without temporal augmentation. In group 1, after splitting the calvarial bone at the diploic space, the inner table was used for covering the bone defect and as an onlay graft for temporal augmentation. The outcome is evaluated by computed tomography at 1-year follow-up. Results: The mean operative time for temporal augmentation was 45 minutes. The mean follow-up was 12 months. The ratio of temporal thickness of operated side to non-operated side was 0.99 in group 1 and 0.44 in group 2, which was statistically different. The mean visual analogue scale score was 1.77 in group 1 and 6.85 in group 2. Conclusion: This study demonstrated a surgical technique using autologous bone graft for successfully preventing the temporal hollowing and improved patient satisfaction.

• Archives of Plastic Surgery
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• v.38 no.1
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• pp.53-62
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• 2011

Purpose: Even though Augmentation rhinoplasty is very popular surgical procedure, it is not easy to obtain ideal materials for augmentation. Many different synthetic materials are used but frequent complications are seen such as infection, extrusion, deform, and dislocation. Autologous tissues were used for augmentation rhinoplasty. We used dermofat graft and fat injection in augmentation rhinoplasty minimizing these problems. Methods: From 2006 to 2009, we used autologous tissues in augmentation rhinoplasty in 40 patients, 20 patients with dermofat graft and other 20 patients were treated with fat injection only. Dermofats were harvested from sacral area. gluteal fold, groin and preexisting scar tissue. Dermofats were inserted with small stab wound and fat tissues were injected as Coleman's technique. The patients were followed up 6 months to 5 years. Results: Most of the patients were satisfied in shape and height the nose. Early complications such as hematoma, infection and seroma were not found. Secondary fat injection was performed in 3 patients (15%) of dermofat graft group instead of 7 patients (35%) of fat injection only group. Conclusion: We obtained satisfactory results in augmentation rhinoplasty with dermofat graft and fat injection. Secondary fat injections were more often in fat injection group than dermofat graft group. Dermofat graft and fat injection could be another alternative technique for augmentation rhinoplasty and fat injection could be a secondary adjunctive treatment for undercorrection due to absorption.

• Archives of Plastic Surgery
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• v.38 no.1
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• pp.15-18
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• 2011

Purpose: The anterior iliac crest is a common source for autologous cancellous bone graft. For patients who have previously received cancellous bone grafts from bilateral anterior iliac crests, there may be concerns of whether a sufficient quantity of autologous cancellous bone remains for additional grafts without harvesting it from other sites, such as the posterior iliac crest. Methods: We experienced 3 cases of reharvesting in 2 patients. The diagnosis of the first patient was bilateral facial cleft number 3. This patient received bilateral side cleft alveoloplasty with corticocancellous bone graft from the both anterior iliac crest respectively by a previous surgeon. This patient then needed reharvesting of the anterior iliac crest cancellous bone to correct an ongoing skeletal problem for the bilateral cleft. The other patient had bilateral incomplete cleft of the primary palate. This patient received left side cleft alveoloplasty with cancellous bone graft from the right anterior iliac crest. Before the patient could receive the alveoloplasty on the other side, a radial head osteotomy and cancellous bone graft was performed by orthopedic surgeons who then used the remaining left iliac crest in order to treat a pulled elbow. For the completion of the right side cleft alveoplasty, the anterior iliac crest cancellous bone needed to be reharvested. Prior to the reharvesting, a preoperative computed tomography scan of the pelvis was obtained to assess the maturity of the donor site regeneration. The grafts were then taken from site where a greater amount of regeneration was evident. Results: Long term follow ups showed that the grafts were successfully taken. This sufficient volume was obtainable 14 months after the first harvest. Conclusion: Satisfactory results were achieved after the reharvesting of iliac cancellous bone. Thus, it appears that the reharvesting of the iliac bone is a possible alternative to multiple site grafting, use of allograft or bone substitute materials.