素輪 善弘

PURPOSE: Triple-negative breast cancer (TNBC) is the most aggressive subtype of breast cancer. Eribulin was approved for the treatment of metastatic breast cancer through the EMBRACE trial, and a subgroup analysis in this clinical trial indicated the efficacy of eribulin in patients with TNBC. However, the prognosis of patients with TNBC is still poor due to various molecular characteristics. Therefore, there is an urgent need for a more effective treatment for the management of TNBC. METHODS: We investigated the synergistic effect of a novel histone deacetylase (HDAC) inhibitor, OBP-801, and eribulin in TNBC cell lines because OBP-801 has been known to enhance the anti-tumor activities of other chemotherapeutic agents. The cell growth was analyzed, and the flow cytometry analysis was conducted to evaluate the effects on cell cycle and the induction of apoptosis. The mechanism underlying the enhancement of inhibition of TNBC cell growth was investigated through Western blot analyses. RESULTS: The combination treatment of OBP-801 with eribulin showed the synergistic inhibition of the growth in TNBC cells, involved with the enhancement of apoptosis. We, for the first time, found that eribulin upregulated survivin and also that OBP-801 could remarkably suppress the upregulation of survivin by eribulin. Moreover, this combination potently suppressed Bcl-xL and the MAPK pathway compared with either agent alone. CONCLUSION: We found that the combination of OBP-801 and eribulin synergistically inhibited the growth with apoptosis in TNBC cells, suggesting that this combination might be a promising novel strategy for treating TNBC patients.

With increasing clinical demands for MEK inhibitors in cancer treatment, overcoming the resistance to MEK inhibitors is an urgent problem to be solved. Numerous reports have shown that MEK inhibition results in the activation of PI3K-Akt signaling, which may confer apoptotic resistance to MEK inhibitors. We here demonstrate that the blockade of the mevalonate pathway using the antilipidemic drug statins represses Akt activation following MEK inhibition and induces significant apoptosis when co-treated with CH5126766 or trametinib. These events were clearly negated by the addition of mevalonate or geranylgeranyl pyrophosphate, indicating that the protein geranylgeranylation is implicated in the apoptotic resistance to MEK inhibitors. Furthermore, mechanistically, the combined treatment of CH5126766 with statins upregulated TNF-related apoptosis-inducing ligand (TRAIL), which was dependent on inhibition of the mevalonate pathway and is involved in apoptosis induction in human breast cancer MDA-MB-231 cells. The present study not only revealed that the mevalonate pathway could be targetable to enhance the efficacy of MEK inhibitors, but also proposes that combinatorial treatment of MEK inhibitors with statins may be a promising therapeutic strategy to sensitize cancer cells to apoptosis.

BACKGROUND: The C-V flap for nipple reconstruction is now one of standard surgical techniques. But decreased projection is still a problem. In recent years, it has been suggested that projection can be more easily maintained when raising of the C-flap is performed with a split thickness dermis. In this study, we examined whether decrease of projection can be prevented by raising of a C-flap with a split dermis rather than with full dermis. METHODS: A total of 49 consecutive patients who underwent reconstruction of a nipple using the C-V flap technique were enrolled. The patients included 22 who underwent surgery using a C-flap with a full thickness dermis (Group F), and 27 who underwent surgery with raising of a flap with a split thickness dermis (Group S). The size of the reconstructed nipple was measured at 2 weeks, 6 months and 1 year postoperatively for comparison between Groups F and S. RESULTS: Partial necrosis of the C-flap end occurred in 4 subjects in only Group S. The decrease in projection after 1 year postoperatively in Group S was significantly lower than that in Group F. In contrast, the teat base size in Group F tended to be greater than that in Group S, suggesting a tendency for an expanded base using a flap with a full dermis. CONCLUSIONS: Our results indicated that it is recommended to use a C-flap with a split dermis for cases with high projection of the nipple on the contralateral side.

Computer-aided design/computer-aided manufacturing (CAD/CAM) guides are now widely used in maxillofacial reconstruction. However, there are few reports of CAD/CAM guides being used for scapular flaps. The authors performed the secondary maxillary and orbital floor reconstruction using a free latissimus dorsi muscle, cutaneous tissue, and scapular flap designed using CAD/CAM techniques in a 72-year-old man who had undergone partial maxillectomy four years previously. The patient had diplopia, the vertical dystopia of eye position, and a large oral-nasal-cutaneous fistula. After the operation, the authors confirmed that the deviation between the postoperative and preoperative planning three-dimensional images was less than 2mm. Because scapular guides require 3 cutting surfaces, the shape of the scapular guide is more complex than that of a conventional fibular guide. In orbital floor reconstruction, the use of a CAM technique such as that used to manufacture the authors' fixation guide is as necessary for accurate, safe, and easy reconstruction as is preoperative CAD planning. The production of a fixation guide as well as a cutting guide is particularly useful because it is difficult to determine the angle for reconstructing the orbital floor by freehand techniques. In this case, the orbital floor was reconstructed based on a mirror image of the healthy side to avoid overcompression of the orbital tissue. Although the patient's vertical dystopia of eye position was improved, diplopia was not improved because, for greater safety, the authors did not plan overcorrection of the orbital volume.

Purpose: The pedicled, descending-branch muscle-sparing latissimus dorsi (MSLD) flap has been widely used for breast reconstruction following total mastectomy. However, the superiority of the MSLD flap compared to the conventional latissimus dorsi (CLD) flap in preventing seroma formation has not been demonstrated. This study compares the morbidities related to seroma formation following pedicled MSLD flap and CLD flap breast reconstruction. Methods: A total of 15 women who underwent partial mastectomy and immediate partial breast reconstruction with MSLD flaps were compared with 15 women under identical conditions with CLD flap breast reconstruction. The medical records were reviewed for both complications and demographic data. The authors compared morbidity, including donor-site seroma, total volume of drain discharge, indwelling period of drainage, and length of hospital stay following both MSLD flap and CLD flap breast reconstruction. Results: The demographic data of the 2 groups were not significantly different. Donor-site seroma occurred in 2 MSLD patients (13.3%) and in 6 CLD patients (40.0%). The total volume of the drain discharge and the indwelling period of drainage at donor site were significantly lower in the MSLD group. The length of hospital stay was significantly shorter (by approximately a day and a half) for the MSLD group. Conclusion: The MSLD flap, with its low complication rate and associated minimal functional and aesthetic deficits at the donor site, may be a useful option for small breast reconstruction if earlier discharge from hospital is demanded.

Background: The optimum number of microvascular anastomoses for safe free tissue transfer is controversial. Although the case for 2 venous anastomoses versus 1 anastomosis has been argued, the use of an additional arterial anastomosis has not been examined in detail. Methods: Twelve patients who underwent 2 arterial anastomoses for a free flap transfer were identified retrospectively from the medical records of patients undergoing reconstruction for head and neck cancer. The free flaps were limited to anterolateral thigh (ALT) flaps. Results: All flaps survived. Complications included venous thrombosis (n = 1), reexploration (n = 1), and leakage (n = 3). The vascular patterns of dual-Arterialized ALT flaps were classified into 3 groups. Types 1 and 2 were ALT flaps that had 2 vascular sources from the descending and lateral branches of the lateral circumflex femoral artery. The number of accompanying veins differed between type 1 (3 veins) and type 2 (2 veins). Type 3 differed from a conventional ALT flap nourished by the descending branch of the lateral circumflex femoral artery (1 vein) by the addition of anastomosis of an artery branching from the descending branch to the vastus medialis muscle. The total operation times for these 3 types of ALT were similar. Conclusions: An additional arterial anastomosis to the free cutaneous flap did not cause any congestion or disturb the balance between inflow and outflow. If the surgeon considers that the first arterial anastomosis is unreliable, an additional anastomosis might be an option in ALT transfer.

Background Computer-aided design and computer-assisted manufacturing (CAD/CAM) of cutting guides are now being used in mandibular reconstruction with fibular bone flaps. Improvements in guide design and accuracy are needed to increase the benefits to patients. Cutting guides have become popular, but fixation guides are rarely considered. Materials and methods The aim of this study was to determine whether using a fixation guide would contribute to better accuracy in mandibular reconstruction. Mandibular segmental osteotomies and fibular reconstructions were performed using model surgery. Models were divided into two groups: without or with a fixation guide (n = 13 for each group). After reconstruction, the distances between reference points such as the condylion laterale (Cl), gonion, and mental tubercle (T) were measured and compared with those of the preoperative virtual plan. Deviations in final positions between the two groups were analyzed. Results The mean deviations were 2.61 mm (range 0.05–7.65 mm) and 2.05 mm (range 0.07–8.52 mm) in the groups without and with a fixation guide, respectively. The overall results were significantly better when a fixation guide was used (p = 0.03). Distances including the Cl reference points such as Cl[sbnd]Cl (p = 0.02) and Cl–T (p = 0.001) were particularly improved. Conclusions Using a fixation guide together with a cutting guide makes mandibular reconstruction more accurate, particularly for positioning the Cl reference points. Therefore, when a CAD/CAM-produced cutting guide is used in preparation for surgery, it is recommended to use a fixation guide as well for more accuracy.

Background Several investigators have evaluated the impaired function of the shoulder after removal of the latissimus dorsi muscle for breast reconstruction. However, a few investigators have studied whether including radiotherapy has a negative effect on functional recovery of the shoulder by a long-term follow-up after surgery. In this study, we compared objective measurements of shoulder function preoperatively and postoperatively for 3 years after latissimus dorsi muscle (LDM) flap transfer and postoperative radiotherapy (PRT). Methods Eighteen patients who underwent unilateral transfer of a pedicled LDM flap and PRT within 2 months of breast-conserving surgery were enrolled in this study. Range of motion (ROM) and muscle strength in exhaustive shoulder movements were measured before surgery, and at 3 and 6 months, and 1 and 3 years. Results The results of ROM measurements at 3months postsurgery showed significant decreases in both flexion and abduction by 7.1 and 9.2 % and at 3 years postsurgery by 4.7 and 5.7 %. The muscle strength measurements at 3 months postsurgery showed statistically significant decreases both in adduction and in the 2nd medial rotation by 30.7 and 25.9 % and at 3 years postsurgery by 36.4 and 20.4 %. A significant improvement in these impairments was not observed for 3 years after surgery compared with that at 3 months after surgery. Conclusion A combination of the LDM flap procedure and PRT could be associated with a higher incidence of tissue adhesions in both flexion and abduction and muscle deficit both in adduction and in the 2nd medial rotation.

Schwann cells (SCs) play pivotal roles in the maintenance and regeneration of the peripheral nervous system. Although transplantation of SCs enhances repair of experimentally damaged peripheral and central nerve tissues, it is difficult to prepare a sufficient number of functional SCs for transplantation therapy without causing adverse events for the donor. Here, we generated functional SCs by somatic cell reprogramming procedures and demonstrated their capability to promote peripheral nerve regeneration. Normal human fibroblasts were phenotypically converted into SCs by transducing SOX10 and Krox20 genes followed by culturing for 10 days resulting in approximately 43% directly converted Schwann cells (dSCs). The dSCs expressed SC-specific proteins, secreted neurotrophic factors, and induced neuronal cells to extend neurites. The dSCs also displayed myelin-forming capability both in vitro and in vivo. Moreover, transplantation of the dSCs into the transected sciatic nerve in mice resulted in significantly accelerated regeneration of the nerve and in improved motor function at a level comparable to that with transplantation of the SCs obtained from a peripheral nerve. The dSCs induced by our procedure may be applicable for novel regeneration therapy for not only peripheral nerve injury but also for central nerve damage and for neurodegenerative disorders related to SC dysfunction.

BACKGROUND: Capsular contracture around implants is a common complication after breast reconstruction. Strain elastography (STE) and shear-wave elastography (SWE) are noninvasive imaging techniques that can measure tissue stiffness and thickness of the capsule. OBJECTIVE: The purposes of the study were to compare STE and SWE for measurement of capsular contracture after breast implant reconstruction using intra-class correlation coefficients (ICCs) and to investigate the correlation of these data with the Baker score, which is the most frequently used clinical staging scale for capsule contracture. METHODS: The subjects were 20 patients (27 implants) who underwent breast reconstruction. RESULTS: The reproducibility of SWE (ICC: 0.878) was higher than that of STE (ICC: 0.724) for measurement of capsular contracture. The correlation coefficient between measurements with the two methods was low (r = 0.6788). The Baker score had a higher correlation with measurements with SWE (r = 0.8124) compared to those with STE (r = 0.6983). CONCLUSIONS: These results suggest that SWE is a better tool for assessment of the degree of capsule contracture surrounding implants after breast reconstruction.

Surgical osteotomy guides created by computer-aided design/computeraided manufacturing (CAD/CAM) have been developed and are now widely used in maxillofacial reconstruction. However, there are no standard procedures for dealing with an intraoperative change in defect size. We report on a case in which we used our CAD/CAM guides to deal with an intraoperative change in defect size in a maxillary reconstruction. We planned the maxillary reconstruction using a free fibula flap because of left maxillary sinus cancer in a 73-year-old man. In Japan, we cannot use commercially supplied CAD/CAM guides because these have not been approved by the government. We created novel CAD/CAM guides by using free software and a low-cost 3D printer. We performed model surgery to check the accuracy of the design and to prebend the titanium plates before the operation. The actual defect in the maxilla was found to be smaller than that used in preoperative planning. It was therefore necessary to rearrange the fibular segments and to rebend the plates. Comparison between the preoperative and postoperative 3D images showed that the deviation was 2-4 mm. In case that the CAD/CAM guides become inapplicable because of an intraoperative change in defect size, rearranging both the ends of set-up fibular segments and rebending the plates in situ allows us to deal with the situation. However, because extra time is needed to rearrange and rebend, the total operation and flap ischemic times are not shortened.

Generation of osteoblasts from human somatic cells may be applicable in an effective transplantation therapy against bone diseases. Recently we established a procedure to directly convert human fibroblasts into osteoblasts by transducing some transcription factor genes via retroviral vectors. However, retroviral vector-mediated transduction may potentially cause tumor formation from the infected cells, thus a non-viral gene transfection method may be more preferable for preparation of osteoblasts to be used for transplantation therapy. Here, we constructed a plasmid vector encoding Oct4, Osterix, and L-Myc that were an appropriate combination of transcription factors for this purpose. Osteoblast-like phenotypes including high alkaline phosphatase (ALP) activity, bone matrix production and osteoblast-specific gene expression were induced in normal human fibroblasts that were transfected with the plasmid followed by culturing in osteogenic medium. The plasmid-driven directly converted osteoblasts (p-dOBs) were obtained even in the absence of a xenogenic protein. The plasmid vector sequence had fallen out of the p-dOBs. The cells formed deposition of calcified bodies in situ after transplantation into mice. These results strongly suggest that p-dOBs can be put into practical use for a novel cell-based therapy against bone diseases. J. Cell. Biochem. 117: 2538-2545, 2016. (c) 2016 Wiley Periodicals, Inc.

Background: During recent decades, multipotent stem cells were found to reside in the adipose tissue, and these adipose-derived stem cells were shown to play beneficial roles, like those of Schwann cells, in peripheral nerve regeneration. However, it has not been well established whether adipose-derived stem cells offer beneficial effects to peripheral nerve injuries in vivo as Schwann cells do. Furthermore, the in situ survival and differentiation of adipose-derived stem cells after transplantation at the injured peripheral nerve tissue remain to be fully elucidated. Methods: Adipose-derived stem cells and Schwann cells were transplanted with gelatin hydrogel tubes at the artificially blunted sciatic nerve lesion in mice. Neuroregenerative abilities of them were comparably estimated. Cre-loxP-mediated fate tracking was performed to visualize survival in vivo of transplanted adipose-derived stem cells and to investigate whether they differentiated into Schwann linage cells at the peripheral nerve injury site. Results: The transplantation of adipose-derived stem cells promoted regeneration of axons, formation of myelin, and restoration of denervation muscle atrophy to levels comparable to those achieved by Schwann cell transplantation. The adipose-derived stem cells survived for at least 4 weeks after transplantation without differentiating into Schwann cells. Conclusions: Transplanted adipose-derived stem cells did not differentiate into Schwann cells but promoted peripheral nerve regeneration at the injured site. The neuroregenerative ability was comparable to that of Schwann cells. Adipose-derived stem cells at an undifferentiated stage may be used as an alternative cell source for autologous cell therapy for patients with peripheral nerve injury.

Background: Intravenous heparin administration is used to prevent thrombosis in free-flap transfer. However, it is unknown whether the use of heparin affects free-flap survival. The purpose of this study is to investigate the effect of heparin in free flap transfer. Methods: Two hundred and six patients who received ablative surgery for head and neck cancer were classified into three groups. Group A received ablative surgery, neck dissection, and free-flap reconstruction, and postoperatively they were administered continuous intravenous unfractionated heparin (5000-10 000 units/day) until postoperative day 7 (POD7); group B received the same procedures as group A but without heparin; group C received only ablative surgery and neck dissection without heparin. As indicators of coagulation time, the prothrombin time-international normalised ratio (PT-INR) and the activated partial thromboplastin time (APTT) were measured, before surgery and on POD1, 3, and 7. Flap failure, bleeding, haematoma formation, re-exploration, and thromboembolic events were recorded. Results: The PT-INR and APTT were 1.3-1.5- times longer in group A (p < 0.01), and 1.3-times longer (p < 0.01) in group B. The PT-INR and APTT were higher in groups A and B than C (p < 0.01). The free-flap success rate was not affected. Only the incidence of haematoma was increased in group A (p = 0.04). Conclusion: Heparin increased the haematoma formation, but did not change the incidence of free-flap failure. Thus, the intravenous low-dose heparin use does not affect microvascular flap survival.

Background: Postoperative seroma is the most common complication of latissimus dorsi (LD) flap surgery for breast reconstruction. The use of EC for elevation of the flap might cause additional risk for seroma formation by injuring surrounding lymph vessels due to heat dispersion. There is a possibility that seroma formation can be prevented by using alternative devices such as harmonic focus (HF) shears that can dissect the tissue simultaneously with sealing the lymph vessels. Methods: Forty-eight patients who underwent breast reconstruction with LD flaps since August 2011 up to April 2015 were enrolled. They were retrospectively split into two groups: 24 in group HF, 24 in group EC (conventional electrocautery). The primary outcome measures were rate of seroma formation and total volume of drain discharge and indwelling period of drainage at the anterior chest and donor site. Secondary outcome measures were length of hospital stay and duration of surgery. Results: The incidence of seroma was 45.8% in the EC group and 20.8% in the HF group. The total volumes of the drain discharge and indwelling period of drainage in the back (donor site) were significantly decreased in the HF group. The length of the hospital stay and surgical time was significantly shorter for the HF group. Conclusions: The use of HF shears on the LD flap donor site is helpful for reducing seroma formation, the length of the drainage period, the surgical time, and the length of the hospital stay.

Fat induration is associated with necrosis and remains a common complication in breast reconstructions with autologous flaps after mastectomy. Fat induration can cause deep tissue infection, pain, distress, and anxiety. However, the diagnosis for this problem has not been objectively defined. In the current article, We will share our experience of using ultrasound shear-wave elastography with 14 patients who had clinical fat induration after breast reconstruction with a deep inferior epigastric perforator (DIEP) flap. The experience suggested that shear wave elastography may be a noninvasive tool to assess alterations of tissue stiffness in a reproducible fashion after breast reconstruction with DIEP flaps. Complications, such as fat necrosis and fatty induration, may occur as a result of unstable blood flow to the flap. Thereby, objective assessments of stiffness might make a major contribution to the understanding of hemodynamics of the DIEP flap after transplantation.

頭頸部癌切除に伴う下顎骨と口腔粘膜の一次再建で,下顎骨区域切除およびチタンプレートと遊離前外側大腿皮弁による再建を行った患者が,その後プレートの破損により下顎骨の偏位と不正咬合をきたしたため,下顎の二次再建を行った.下顎骨区域切除による骨欠損があるため,整復位置を正確に決めることが困難であると考えられたので,三次元実体モデルを用いた術前シミュレーション手術を行い,術中整復位をあらかじめ設定した.手術シミュレーションにより,1,下顎の不正咬合の現状把握や,整復位での残存下顎骨の位置確認が明瞭となった.2,下顎骨の整復位を保つメモリープレート,あるいは移植腓骨を固定するチタンプレートの,術前プレート屈曲,テンプレート作成による手術時間短縮が可能であった.3,下顎骨欠損に充填する腓骨の骨切り部位と形状の術前予測による模擬手術ができ,手術時間短縮および視覚化によるより簡潔で正確な手術操作への転換ができた.などの利点があった.欠点は,術前に準備時間がかかること,保険適応ではあるが経済的な問題から,モデル作成作業を医師が行わなくてはいけないこと,などであった.形成外科・頭頸部再建領域では三次元実体モデルを用いた手術支援はまれであるため,その経験と特徴について報告した.(著者抄録)